LOCUS       BB3_pGAP_Linker_BbsI(intern)_pPFK300_Cas9_cycTT_NTC      8089 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Fragment        1..678
                     /origin="pSTBLUE-1 wo_ORI_circular_pUC_ORI_aus_pGAPZB
                     (2.745bp circular)Fragment: from BpiI cut at 2054^2055 to
                     BpiI cut at 2732^2733, 678bpOrientation: originalPlus
                     strand 5':   no changePlus strand 3':   no changeMinus
                     strand 5':  no changeMinus strand 3':  no change"
     Vector          join(6446..8089,1..722)
                     /origin="cloned empty BB3_ARSCEN_NTC_AC (Franz) new
                     (2,396bp circular)Target vector: from BsaI cut at 752^753
                     to BsaI cut at 722^723, 2,366bpOrientation: originalPlus
                     strand 5':   no changePlus strand 3':   no changeMinus
                     strand 5':  no changeMinus strand 3':  no change"
     oriT            complement(5..678)
                     /label="pUC_ORI_aus_pGAPZB"
     primer          complement(53..71)
                     /label="B1_08"
     primer          440..459
                     /label="J_77"
     Fragment        join(<6446..6449,679..>722)
                     /origin="Linker_FS1FsA_BX_FsCFS2_FW (106bp
                     linear)Fragment: from BpiI cut at 12^13 to BpiI cut at
                     90^91, 78bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     terminator      683..722
                     /label="BBa_B1007"
     FS              723..726
                     /label="FS A"
     Vector          join(1743..>1746,<723..726)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 726^727
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        729..>1221
                     /label="pGAP"
     Restriction_sit join(1221^1222,complement(1221^1222))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1222..>1253
                     /label="L_FS23_BbsI_fw"
     Linker          1222..1253
                     /origin="PCR_Linker_FS23_BbsI(intern) (54bp
                     linear)Fragment: from BpiI cut at 9^10 to BpiI cut at
                     41^42, 32bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="FS 23 Linker"
     FS              1222..1225
                     /label="FS2"
     primer          complement(<1226..>1257)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1254..1286,complement(1287..1719),complement(1720..>1746))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1258..>1742
                     /note="Forward primer name: RPS25ATT_M1_FW"
                     /note="Forward primer: GacagGtcTccgCTTCCTTGTCGAGTGTGTTT"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_RV"
                     /note="Reverse primer:
                     gatcggtctCcAgctGTCAAATATTCAGATGGTGA"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 458"
                     /label="rps25aTT"
     FS              1743..1746
                     /label="FS 4"
     primer          <1747..>1750
                     /label="3_BsaI_FsB_Bam"
     misc_feature    join(<6442..>6445,<1747..>1750)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (108bp linear)Fragment: full
                     sequenceOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1747..1750
                     /label="FS B"
     Vector          join(6442..>6445,<1747..1750)
                     /origin="BB2_L_BC_syn_BbsI_7 (2,215bp circular)Target
                     vector: from BpiI cut at 639^640 to BpiI cut at 607^608,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          complement(<1751..>1754)
                     /label="4_BsaI_FsB_BamXho"
     promoter        1763..1978
                     /label="pPpPfk"
     primer          <1979..2003
                     /label="FS2_Cas9_BsaI_fw"
     CDS             1980..6119
                     /label="Cas9"
     Taming          2030
                     /label="C>T"
     primer          complement(3391..3410)
                     /label="1_cas9_rev"
     primer          3391..3410
                     /label="1_cas9_fw"
     Taming          3395
                     /label="C>t"
     primer          complement(3665..3686)
                     /label="2_cas9_rev"
     primer          3665..3686
                     /label="2_cas9_fw"
     Taming          3677
                     /label="A>g"
     primer          complement(4134..4151)
                     /label="3_cas9_rev"
     primer          4134..4151
                     /label="3_cas9_fw"
     Taming          4136
                     /label="T>c"
     primer          complement(4849..4870)
                     /label="4_cas9_rev"
     primer          4849..4870
                     /label="4_cas9_fw"
     Taming          4865
                     /label="G>c"
     primer          complement(5716..5733)
                     /label="5_cas9_rev"
     primer          5716..5734
                     /label="5_cas9_fw"
     Taming          5723
                     /label="T>c"
     NLS             6084..6119
                     /label="NLS SV40"
     primer          complement(6097..>6123)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6120..>6441
                     /label="FS3_CYCtt_FS4"
     primer          <6442..>6445
                     /label="5_BsaI_FsC_Xho"
     FS              6442..6445
                     /label="FS 4"
     primer          complement(6446..>6449)
                     /label="6_BsaI_FsC"
     primer          <6450..6468
                     /label="ntcMX frag1 fwd"
     Fragment        6450..7573
                     /origin="ntcMX frag (1.150bp linear)Fragment: from BpiI
                     cut at 14^15 to BpiI cut at 1138^1139,
                     1.124bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     promoter        6454..6792
                     /label="A. gossipii TEFprom"
     primer          complement(6681..6700)
                     /label="J_78"
     CDS             6798..7372
                     /label="nat1"
     mutation        6902
                     /label="C/T"
     mutation        6905
                     /label="C/T"
     terminator      7377..7573
                     /label="A. gossipii TEFterm"
     primer          complement(7556..>7577)
                     /label="ntcMX frag2 rev"
     misc_feature    <7574..8089
                     /label="CEN6_ARS4"
     primer          <7574..7592
                     /label="CEN6/ARS4_fwd"
     primer          7574..7592
                     /label="CEN6/ARS4_fwd_partial"
     Fragment        7574..8089
                     /origin="CEN6_ARS4_flanked_FS3_FS4_new (with primers)
                     (544bp linear)Fragment: from BpiI cut at 12^13 to BpiI
                     cut at 528^529, 516bpOrientation: originalPlus strand 5':
                     no changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     modified_base   7799
                     /label="G/A"
     primer          complement(8073..8089)
                     /label="CEN6/ARS4_rev_partial"
     primer          complement(8073..8089)
                     /label="CEN6/ARS4_rev"
ORIGIN
        1 CGCTGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT
       61 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT
      121 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC
      181 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA
      241 GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT
      301 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA
      361 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG
      421 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC
      481 CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA
      541 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG
      601 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT
      661 TGATCTTTTC TACGGGGTGG AGcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcGATCGGAG GATCCTTTTT TGTAGAAATG TCTTGGTGTC CTCGTCCAAT CAGGTAGCCA
      781 TCTCTGAAAT ATCTGGCTCC GTTGCAACTC CGAACGACCT GCTGGCAACG TAAAATTCTC
      841 CGGGGTAAAA CTTAAATGTG GAGTAATGGA ACCAGAAACG TCTCTTCCCT TCTCTCTCCT
      901 TCCACCGCCC GTTACCGTCC CTAGGAAATT TTACTCTGCT GGAGAGCTTC TTCTACGGCC
      961 CCCTTGCAGC AATGCTCTTC CCAGCATTAC GTTGCGGGTA AAACGGAGGT CGTGTACCCG
     1021 ACCTAGCAGC CCAGGGATGG AAAAGTCCCG GCCGTCGCTG GCAATAATAG CGGGCGGACG
     1081 CATGTCATGA GATTATTGGA AACCACCAGA ATCGAATATA AAAGGCGAAC ACCTTTCCCA
     1141 ATTTTGGTTT CTCCTGACCC AAAGACTTTA AATTTAATTT ATTTGTCCCT ATTTCAATCA
     1201 ATTGAACAAC TATCAAAACA CCATGGCGTC TTCGGATCCC TCGAGGAAGA CGCGCTTTTA
     1261 GTGTACATCT GATAATATAG TACTACCACG TATGATAATG TAGAGAATAG GCTTCCTTGT
     1321 CGAGTGTGTT TGCAGTTTTC TTGAGTTTCA AGGTTTAAAT GCTGGTATAT TAGTTCATCG
     1381 AAGGTTTCAG CCAATAGCAC CTTAAATCAA TCAAACTAAT TCGACTCTTA CGAAAGAGCC
     1441 TACTGTGTTT AGTATCGAAG TCGTTTACCT TTCATGTTGA ATAGCTTCCT CTCTGACCCT
     1501 AACATTTCAA GATCCTCCTA AAGTTACCCG GATTGTGAAA TTCTAATGAT CCACCTGCCC
     1561 AATGCATTTT TTCTTTATTC AGTTTACCTT TTTTACCTAA TATACGAGCT TGTTAAAGTA
     1621 AGTGGCACTG CAATACTAGG CTTATTGTTG ATATTATGAT GAATCGTTTT CACAAACTTG
     1681 ATTTCCTGTG AACTCACCAT GTACTAAGGA AAAAAACATG CATCACCATC TGAATATTTG
     1741 ACCGCTCCGG GGAGCCTGCC ATTAGGCAAA CACCAACTAG GAACCTACAC CAAGTTATGA
     1801 AGGAGCTTGG AAAGGATTAT AAAAAATGCG ATTACCTAAT ACACACACGA TGCCTGAGCG
     1861 GTTAATGTGA AATTTCCCGG CCATCGATCG CCCCACAATC AGACTATTTT CGCTCATACA
     1921 ATAAAAAGTC TCCTGTTTCC ACCCGAGTAT CACCAGAACA AAGTAACAAA TACAAAACCA
     1981 TGGACAAGAA GTACTCCATT GGGCTCGATA TCGGCACAAA CAGCGTCGGT TGGGCCGTCA
     2041 TTACGGACGA GTACAAGGTG CCGAGCAAAA AATTCAAAGT TCTGGGCAAT ACCGATCGCC
     2101 ACAGCATAAA GAAGAACCTC ATTGGCGCCC TCCTGTTCGA CTCCGGGGAG ACGGCCGAAG
     2161 CCACGCGGCT CAAAAGAACA GCACGGCGCA GATATACCCG CAGAAAGAAT CGGATCTGCT
     2221 ACCTGCAGGA GATCTTTAGT AATGAGATGG CTAAGGTGGA TGACTCTTTC TTCCATAGGC
     2281 TGGAGGAGTC CTTTTTGGTG GAGGAGGATA AAAAGCACGA GCGCCACCCA ATCTTTGGCA
     2341 ATATCGTGGA CGAGGTGGCG TACCATGAAA AGTACCCAAC CATATATCAT CTGAGGAAGA
     2401 AGCTTGTAGA CAGTACTGAT AAGGCTGACT TGCGGTTGAT CTATCTCGCG CTGGCGCATA
     2461 TGATCAAATT TCGGGGACAC TTCCTCATCG AGGGGGACCT GAACCCAGAC AACAGCGATG
     2521 TCGACAAACT CTTTATCCAA CTGGTTCAGA CTTACAATCA GCTTTTCGAA GAGAACCCGA
     2581 TCAACGCATC CGGAGTTGAC GCCAAAGCAA TCCTGAGCGC TAGGCTGTCC AAATCCCGGC
     2641 GGCTCGAAAA CCTCATCGCA CAGCTCCCTG GGGAGAAGAA GAACGGCCTG TTTGGTAATC
     2701 TTATCGCCCT GTCACTCGGG CTGACCCCCA ACTTTAAATC TAACTTCGAC CTGGCCGAAG
     2761 ATGCCAAGCT TCAACTGAGC AAAGACACCT ACGATGATGA TCTCGACAAT CTGCTGGCCC
     2821 AGATCGGCGA CCAGTACGCA GACCTTTTTT TGGCGGCAAA GAACCTGTCA GACGCCATTC
     2881 TGCTGAGTGA TATTCTGCGA GTGAACACGG AGATCACCAA AGCTCCGCTG AGCGCTAGTA
     2941 TGATCAAGCG CTATGATGAG CACCACCAAG ACTTGACTTT GCTGAAGGCC CTTGTCAGAC
     3001 AGCAACTGCC TGAGAAGTAC AAGGAAATTT TCTTCGATCA GTCTAAAAAT GGCTACGCCG
     3061 GATACATTGA CGGCGGAGCA AGCCAGGAGG AATTTTACAA ATTTATTAAG CCCATCTTGG
     3121 AAAAAATGGA CGGCACCGAG GAGCTGCTGG TAAAGCTTAA CAGAGAAGAT CTGTTGCGCA
     3181 AACAGCGCAC TTTCGACAAT GGAAGCATCC CCCACCAGAT TCACCTGGGC GAACTGCACG
     3241 CTATCCTCAG GCGGCAAGAG GATTTCTACC CCTTTTTGAA AGATAACAGG GAAAAGATTG
     3301 AGAAAATCCT CACATTTCGG ATACCCTACT ATGTAGGCCC CCTCGCCCGG GGAAATTCCA
     3361 GATTCGCGTG GATGACTCGC AAATCAGAAG AGACtATCAC TCCCTGGAAC TTCGAGGAAG
     3421 TCGTGGATAA GGGGGCCTCT GCCCAGTCCT TCATCGAAAG GATGACTAAC TTTGATAAAA
     3481 ATCTGCCTAA CGAAAAGGTG CTTCCTAAAC ACTCTCTGCT GTACGAGTAC TTCACAGTTT
     3541 ATAACGAGCT CACCAAGGTC AAATACGTCA CAGAAGGGAT GAGAAAGCCA GCATTCCTGT
     3601 CTGGAGAGCA GAAGAAAGCT ATCGTGGACC TCCTCTTCAA GACGAACCGG AAAGTTACCG
     3661 TGAAACAGCT CAAAGAgGAC TATTTCAAAA AGATTGAATG TTTCGACTCT GTTGAAATCA
     3721 GCGGAGTGGA GGATCGCTTC AACGCATCCC TGGGAACGTA TCACGATCTC CTGAAAATCA
     3781 TTAAAGACAA GGACTTCCTG GACAATGAGG AGAACGAGGA CATTCTTGAG GACATTGTCC
     3841 TCACCCTTAC GTTGTTTGAA GATAGGGAGA TGATTGAAGA ACGCTTGAAA ACTTACGCTC
     3901 ATCTCTTCGA CGACAAAGTC ATGAAACAGC TCAAGAGGCG CCGATATACA GGATGGGGGC
     3961 GGCTGTCAAG AAAACTGATC AATGGGATCC GAGACAAGCA GAGTGGAAAG ACAATCCTGG
     4021 ATTTTCTTAA GTCCGATGGA TTTGCCAACC GGAACTTCAT GCAGTTGATC CATGATGACT
     4081 CTCTCACCTT TAAGGAGGAC ATCCAGAAAG CACAAGTTTC TGGCCAGGGG GACAGcCTTC
     4141 ACGAGCACAT CGCTAATCTT GCAGGTAGCC CAGCTATCAA AAAGGGAATA CTGCAGACCG
     4201 TTAAGGTCGT GGATGAACTC GTCAAAGTAA TGGGAAGGCA TAAGCCCGAG AATATCGTTA
     4261 TCGAGATGGC CCGAGAGAAC CAAACTACCC AGAAGGGACA GAAGAACAGT AGGGAAAGGA
     4321 TGAAGAGGAT TGAAGAGGGT ATAAAAGAAC TGGGGTCCCA AATCCTTAAG GAACACCCAG
     4381 TTGAAAACAC CCAGCTTCAG AATGAGAAGC TCTACCTGTA CTACCTGCAG AACGGCAGGG
     4441 ACATGTACGT GGATCAGGAA CTGGACATCA ATCGGCTCTC CGACTACGAC GTGGATCATA
     4501 TCGTGCCCCA GTCTTTTCTC AAAGATGATT CTATTGATAA TAAAGTGTTG ACAAGATCCG
     4561 ATAAAAATAG AGGGAAGAGT GATAACGTCC CCTCAGAAGA AGTTGTCAAG AAAATGAAAA
     4621 ATTATTGGCG GCAGCTGCTG AACGCCAAAC TGATCACACA ACGGAAGTTC GATAATCTGA
     4681 CTAAGGCTGA ACGAGGTGGC CTGTCTGAGT TGGATAAAGC CGGCTTCATC AAAAGGCAGC
     4741 TTGTTGAGAC ACGCCAGATC ACCAAGCACG TGGCCCAAAT TCTCGATTCA CGCATGAACA
     4801 CCAAGTACGA TGAAAATGAC AAACTGATTC GAGAGGTGAA AGTTATTACT CTGAAGTCTA
     4861 AGCTcGTCTC AGATTTCAGA AAGGACTTTC AGTTTTATAA GGTGAGAGAG ATCAACAATT
     4921 ACCACCATGC GCATGATGCC TACCTGAATG CAGTGGTAGG CACTGCACTT ATCAAAAAAT
     4981 ATCCCAAGCT TGAATCTGAA TTTGTTTACG GAGACTATAA AGTGTACGAT GTTAGGAAAA
     5041 TGATCGCAAA GTCTGAGCAG GAAATAGGCA AGGCCACCGC TAAGTACTTC TTTTACAGCA
     5101 ATATTATGAA TTTTTTCAAG ACCGAGATTA CACTGGCCAA TGGAGAGATT CGGAAGCGAC
     5161 CACTTATCGA AACAAACGGA GAAACAGGAG AAATCGTGTG GGACAAGGGT AGGGATTTCG
     5221 CGACAGTCCG GAAGGTCCTG TCCATGCCGC AGGTGAACAT CGTTAAAAAG ACCGAAGTAC
     5281 AGACCGGAGG CTTCTCCAAG GAAAGTATCC TCCCGAAAAG GAACAGCGAC AAGCTGATCG
     5341 CACGCAAAAA AGATTGGGAC CCCAAGAAAT ACGGCGGATT CGATTCTCCT ACAGTCGCTT
     5401 ACAGTGTACT GGTTGTGGCC AAAGTGGAGA AAGGGAAGTC TAAAAAACTC AAAAGCGTCA
     5461 AGGAACTGCT GGGCATCACA ATCATGGAGC GATCAAGCTT CGAAAAAAAC CCCATCGACT
     5521 TTCTCGAGGC GAAAGGATAT AAAGAGGTCA AAAAAGACCT CATCATTAAG CTTCCCAAGT
     5581 ACTCTCTCTT TGAGCTTGAA AACGGCCGGA AACGAATGCT CGCTAGTGCG GGCGAGCTGC
     5641 AGAAAGGTAA CGAGCTGGCA CTGCCCTCTA AATACGTTAA TTTCTTGTAT CTGGCCAGCC
     5701 ACTATGAAAA GCTCAAAGGG TCcCCCGAAG ATAATGAGCA GAAGCAGCTG TTCGTGGAAC
     5761 AACACAAACA CTACCTTGAT GAGATCATCG AGCAAATAAG CGAATTCTCC AAAAGAGTGA
     5821 TCCTCGCCGA CGCTAACCTC GATAAGGTGC TTTCTGCTTA CAATAAGCAC AGGGATAAGC
     5881 CCATCAGGGA GCAGGCAGAA AACATTATCC ACTTGTTTAC TCTGACCAAC TTGGGCGCGC
     5941 CTGCAGCCTT CAAGTACTTC GACACCACCA TAGACAGAAA GCGGTACACC TCTACAAAGG
     6001 AGGTCCTGGA CGCCACACTG ATTCATCAGT CAATTACGGG GCTCTATGAA ACAAGAATCG
     6061 ACCTCTCTCA GCTCGGTGGA GACAGCAGGG CTGACCCCAA GAAGAAGAGG AAGGTGTGAG
     6121 CTTCGTCCGA CGGCGGCCCA CGGGTCCCAG GCCTCGGAGA TCCGTCCCCC TTTTCCTTTG
     6181 TCGATATCAT GTAATTAGTT ATGTCACGCT TACATTCACG CCCTCCCCCC ACATCCGCTC
     6241 TAACCGAAAA GGAAGGAGTT AGACAACCTG AAGTCTAGGT CCCTATTTAT TTTTTTATAG
     6301 TTATGTTAGT ATTAAGAACG TTATTTATAT TTCAAATTTT TCTTTTTTTT CTGTACAGAC
     6361 GCGTGTACGC ATGTAACATT ATACTGAAAA CCTTGCTTGA GAAGGTTTTG GGACGCTCGA
     6421 AGGCTTTAAT TTGCAAGCTG GCGCTAATTC ATGGACATGG AGGCCCAGAA TACCCTCCTT
     6481 GACAGTCTTG ACGTGCGCAG CTCAGGGGCA TGATGTGACT GTCGCCCGTA CATTTAGCCC
     6541 ATACATCCCC ATGTATAATC ATTTGCATCC ATACATTTTG ATGGCCGCAC GGCGCGAAGC
     6601 AAAAATTACG GCTCCTCGCT GCAGACCTGC GAGCAGGGAA ACGCTCCCCT CACAGACGCG
     6661 TTGAATTGTC CCCACGCCGC GCCCCTGTAG AGAAATATAA AAGGTTAGGA TTTGCCACTG
     6721 AGGTTCTTCT TTCATATACT TCCTTTTAAA ATCTTGCTAG GATACAGTTC TCACATCACA
     6781 TCCGAACATA AACAACCATG ggtaccactc ttgacgacac ggcttaccgg taccgcacca
     6841 gtgtcccggg ggacgccgag gccatcgagg cactggatgg gtccttcacc accgacaccg
     6901 tTTTTcgcgt caccgccacc ggggacggct tcaccctgcg ggaggtgccg gtggacccgc
     6961 ccctgaccaa ggtgttcccc gacgacgaat cggacgacga atcggacgac ggggaggacg
     7021 gcgacccgga ctcccggacg ttcgtcgcgt acggggacga cggcgacctg gcgggcttcg
     7081 tggtcgtctc gtactccggc tggaaccgcc ggctgaccgt cgaggacatc gaggtcgccc
     7141 cggagcaccg ggggcacggg gtcgggcgcg cgttgatggg gctcgcgacg gagttcgccc
     7201 gcgagcgggg cgccgggcac ctctggctgg aggtcaccaa cgtcaacgca ccggcgatcc
     7261 acgcgtaccg gcggatgggg ttcaccctct gcggcctgga caccgccctg tacgacggca
     7321 ccgcctcgga cggcgagcag gcgctctaca tgagcatgcc ctgcccctaa tcagtACTGA
     7381 CAATAAAAAG ATTCTTGTTT TCAAGAACTT GTCATTTGTA TAGTTTTTTT ATATTGTAGT
     7441 TGTTCTATTT TAATCAAATG TTAGCGTGAT TTATATTTTT TTTCGCCTCG ACATCATCTG
     7501 CCCAGATGCG AAGTTAAGTG CGCAGAAAGT AATATCATGC GTCAATCGTA TGTGAATGCT
     7561 GGTCGCTATA CTGGCTTCCT TTTCATCACG TGCTATAAAA ATAATTATAA TTTAAATTTT
     7621 TTAATATAAA TATATAAATT AAAAATAGAA AGTAAAAAAA GAAATTAAAG AAAAAATAGT
     7681 TTTTGTTTTC CGAAGATGTA AAAGACTCTA GGGGGATCGC CAACAAATAC TACCTTTTAT
     7741 CTTGCTCTTC CTGCTCTCAG GTATTAATGC CGAATTGTTT CATCTTGTCT GTGTAGAACA
     7801 CCACACACGA AAATCCTGTG ATTTTACATT TTACTTATCG TTAATCGAAT GTATATCTAT
     7861 TTAATCTGCT TTTCTTGTCT AATAAATATA TATGTAAAGT ACGCTTTTTG TTGAAATTTT
     7921 TTAAACCTTT GTTTATTTTT TTTTCTTCAT TCCGTAACTC TTCTACCTTC TTTATTTACT
     7981 TTCTAAAATC CAAATACAAA ACATAAAAAT AAATAAACAC AGAGTAAATT CCCAAATTAT
     8041 TCCATCATTA AAAGATACGA GGCGCGTGTA AGTTACAGGC AAGCGATCG
//
LOCUS       BB3_pGAP_Linker_BbsI(intern)_RPS25Att_pLAT1_Cas9_cycTT_NTC      8866 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Fragment        1..678
                     /origin="pSTBLUE-1 wo_ORI_circular_pUC_ORI_aus_pGAPZB
                     (2.745bp circular)Fragment: from BpiI cut at 2054^2055 to
                     BpiI cut at 2732^2733, 678bpOrientation: originalPlus
                     strand 5':   no changePlus strand 3':   no changeMinus
                     strand 5':  no changeMinus strand 3':  no change"
     Vector          join(7223..8866,1..722)
                     /origin="cloned empty BB3_ARSCEN_NTC_AC (Franz) new
                     (2,396bp circular)Target vector: from BsaI cut at 752^753
                     to BsaI cut at 722^723, 2,366bpOrientation: originalPlus
                     strand 5':   no changePlus strand 3':   no changeMinus
                     strand 5':  no changeMinus strand 3':  no change"
     oriT            complement(5..678)
                     /label="pUC_ORI_aus_pGAPZB"
     primer          complement(53..71)
                     /label="B1_08"
     primer          440..459
                     /label="J_77"
     Fragment        join(<7223..7226,679..>722)
                     /origin="Linker_FS1FsA_BX_FsCFS2_FW (106bp
                     linear)Fragment: from BpiI cut at 12^13 to BpiI cut at
                     90^91, 78bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     terminator      683..722
                     /label="BBa_B1007"
     FS              723..726
                     /label="FS A"
     Vector          join(1743..>1746,<723..726)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 726^727
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        729..>1221
                     /label="pGAP"
     Restriction_sit join(1221^1222,complement(1221^1222))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1222..>1253
                     /label="L_FS23_BbsI_fw"
     Linker          1222..1253
                     /origin="PCR_Linker_FS23_BbsI(intern) (54bp
                     linear)Fragment: from BpiI cut at 9^10 to BpiI cut at
                     41^42, 32bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="FS 23 Linker BpiI"
     FS              1222..1225
                     /label="FS2"
     primer          complement(<1226..>1257)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1254..1286,complement(1287..1719),complement(1720..>1746))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1258..>1742
                     /note="Forward primer name: RPS25ATT_M1_FW"
                     /note="Forward primer: GacagGtcTccgCTTCCTTGTCGAGTGTGTTT"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_RV"
                     /note="Reverse primer:
                     gatcggtctCcAgctGTCAAATATTCAGATGGTGA"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 458"
                     /label="rps25aTT"
     FS              1743..1746
                     /label="FS 4"
     primer          <1747..>1750
                     /label="3_BsaI_FsB_Bam"
     misc_feature    join(<7219..>7222,<1747..>1750)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (108bp linear)Fragment: full
                     sequenceOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1747..1750
                     /label="FS B"
     Vector          join(7219..>7222,<1747..1750)
                     /origin="BB2_L_BC_syn_BbsI_7 (2,215bp circular)Target
                     vector: from BpiI cut at 639^640 to BpiI cut at 607^608,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          complement(<1751..>1754)
                     /label="4_BsaI_FsB_BamXho"
     promoter        <1751..>2755
                     /locus_tag="PAS_chr1-1_0049"
                     /codon_start=1
                     /product="Ribonuclease H2 catalytic subunit, removes RNA
                     primers during Okazaki fragment synthesis"
                     /protein_id="XP_002489688.1"
                     /db_xref="GI:254565155"
                     /db_xref="GeneID:8197566"
                     /translation="MTRIENRVCDSSPMTTTFIPPSVPEISTYRTTTYVSEIPESVLK
                     NPEKPCCLGVDEAGRGPVVGPMVYGVSYCLEEYEPRLKDFGFADSKTLTEQKRSELLE
                     SLCNGDHELRKNVGWATTSISACDISSAMLKPHNIGNYNLNEQAHDVTMELIDKVIQK
                     GVNLRHVYVDTVGPPASYQKKLQQRFPALLITVSKKADAIYPCVSSASVCAKVTRDVC
                     LRTSNGEENLLDCGSGYPSDPNTKKWLAQSIHPLFGWSRDVRFSWQTAKDCLERHNSI
                     KVVFGEEEPTPVNFGDLGLSKQKTDFKDLEISTSWWGH"
                     /label="pLAT1"
     Site            join(2755^2756,complement(2755^2756),complement(2755^2756))
                     /site_type="restriction site"
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 3162^3163"
                     /note="cut_0_on_negative_strand: 3166^3167"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <2756..2780
                     /label="FS2_Cas9_BsaI_fw"
     CDS             2757..6896
                     /label="Cas9"
     Taming          2807
                     /label="C>T"
     primer          complement(4168..4187)
                     /label="1_cas9_rev"
     primer          4168..4187
                     /label="1_cas9_fw"
     Taming          4172
                     /label="C>t"
     primer          complement(4442..4463)
                     /label="2_cas9_rev"
     primer          4442..4463
                     /label="2_cas9_fw"
     Taming          4454
                     /label="A>g"
     primer          complement(4911..4928)
                     /label="3_cas9_rev"
     primer          4911..4928
                     /label="3_cas9_fw"
     Taming          4913
                     /label="T>c"
     primer          complement(5626..5647)
                     /label="4_cas9_rev"
     primer          5626..5647
                     /label="4_cas9_fw"
     Taming          5642
                     /label="G>c"
     primer          complement(6493..6510)
                     /label="5_cas9_rev"
     primer          6493..6511
                     /label="5_cas9_fw"
     Taming          6500
                     /label="T>c"
     NLS             6861..6896
                     /label="NLS SV40"
     primer          complement(6874..>6900)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6897..>7218
                     /label="FS3_CYCtt_FS4"
     primer          <7219..>7222
                     /label="5_BsaI_FsC_Xho"
     FS              7219..7222
                     /label="FS 4"
     primer          complement(7223..>7226)
                     /label="6_BsaI_FsC"
     primer          <7227..7245
                     /label="ntcMX frag1 fwd"
     Fragment        7227..8350
                     /origin="ntcMX frag (1.150bp linear)Fragment: from BpiI
                     cut at 14^15 to BpiI cut at 1138^1139,
                     1.124bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     promoter        7231..7569
                     /label="A. gossipii TEFprom"
     primer          complement(7458..7477)
                     /label="J_78"
     CDS             7575..8149
                     /label="nat1"
     mutation        7679
                     /label="C/T"
     mutation        7682
                     /label="C/T"
     terminator      8154..8350
                     /label="A. gossipii TEFterm"
     primer          complement(8333..>8354)
                     /label="ntcMX frag2 rev"
     misc_feature    <8351..8866
                     /label="CEN6_ARS4"
     primer          <8351..8369
                     /label="CEN6/ARS4_fwd"
     primer          8351..8369
                     /label="CEN6/ARS4_fwd_partial"
     Fragment        8351..8866
                     /origin="CEN6_ARS4_flanked_FS3_FS4_new (with primers)
                     (544bp linear)Fragment: from BpiI cut at 12^13 to BpiI
                     cut at 528^529, 516bpOrientation: originalPlus strand 5':
                     no changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     modified_base   8576
                     /label="G/A"
     primer          complement(8850..8866)
                     /label="CEN6/ARS4_rev_partial"
     primer          complement(8850..8866)
                     /label="CEN6/ARS4_rev"
ORIGIN
        1 CGCTGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT
       61 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT
      121 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC
      181 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA
      241 GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT
      301 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA
      361 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG
      421 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC
      481 CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA
      541 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG
      601 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT
      661 TGATCTTTTC TACGGGGTGG AGcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcGATCGGAG GATCCTTTTT TGTAGAAATG TCTTGGTGTC CTCGTCCAAT CAGGTAGCCA
      781 TCTCTGAAAT ATCTGGCTCC GTTGCAACTC CGAACGACCT GCTGGCAACG TAAAATTCTC
      841 CGGGGTAAAA CTTAAATGTG GAGTAATGGA ACCAGAAACG TCTCTTCCCT TCTCTCTCCT
      901 TCCACCGCCC GTTACCGTCC CTAGGAAATT TTACTCTGCT GGAGAGCTTC TTCTACGGCC
      961 CCCTTGCAGC AATGCTCTTC CCAGCATTAC GTTGCGGGTA AAACGGAGGT CGTGTACCCG
     1021 ACCTAGCAGC CCAGGGATGG AAAAGTCCCG GCCGTCGCTG GCAATAATAG CGGGCGGACG
     1081 CATGTCATGA GATTATTGGA AACCACCAGA ATCGAATATA AAAGGCGAAC ACCTTTCCCA
     1141 ATTTTGGTTT CTCCTGACCC AAAGACTTTA AATTTAATTT ATTTGTCCCT ATTTCAATCA
     1201 ATTGAACAAC TATCAAAACA CCATGGCGTC TTCGGATCCC TCGAGGAAGA CGCGCTTTTA
     1261 GTGTACATCT GATAATATAG TACTACCACG TATGATAATG TAGAGAATAG GCTTCCTTGT
     1321 CGAGTGTGTT TGCAGTTTTC TTGAGTTTCA AGGTTTAAAT GCTGGTATAT TAGTTCATCG
     1381 AAGGTTTCAG CCAATAGCAC CTTAAATCAA TCAAACTAAT TCGACTCTTA CGAAAGAGCC
     1441 TACTGTGTTT AGTATCGAAG TCGTTTACCT TTCATGTTGA ATAGCTTCCT CTCTGACCCT
     1501 AACATTTCAA GATCCTCCTA AAGTTACCCG GATTGTGAAA TTCTAATGAT CCACCTGCCC
     1561 AATGCATTTT TTCTTTATTC AGTTTACCTT TTTTACCTAA TATACGAGCT TGTTAAAGTA
     1621 AGTGGCACTG CAATACTAGG CTTATTGTTG ATATTATGAT GAATCGTTTT CACAAACTTG
     1681 ATTTCCTGTG AACTCACCAT GTACTAAGGA AAAAAACATG CATCACCATC TGAATATTTG
     1741 ACCGCTCCGG GGAGGGAAGA ATATGAACCA CGATTGAAAG ACTTTGGGTT TGCCGATTCT
     1801 AAAACACTTA CTGAACAGAA AAGAAGTGAA TTATTAGAGA GTTTATGCAA CGGAGATCAT
     1861 GAGCTTAGAA AGAACGTAGG CTGGGCAACA ACATCCATTT CTGCCTGCGA TATTTCCAGT
     1921 GCAATGCTAA AGCCACATAA TATAGGCAAT TACAACTTGA ATGAGCAGGC TCATGATGTC
     1981 ACCATGGAAC TGATAGATAA AGTCATCCAA AAAGGTGTCA ATTTACGGCA CGTCTACGTA
     2041 GATACTGTCG GACCTCCAGC ATCATACCAA AAGAAACTGC AACAAAGGTT TCCCGCACTG
     2101 CTGATCACCG TATCTAAAAA AGCAGATGCC ATCTATCCAT GTGTTTCAAG TGCTTCAGTT
     2161 TGCGCCAAGG TCACTAGAGA CGTCTGTTTG AGAACCTCAA ATGGGGAGGA GAACCTATTA
     2221 GATTGTGGGT CTGGATATCC CTCGGACCCA AATACTAAGA AATGGCTAGC CCAGTCTATA
     2281 CATCCATTGT TTGGCTGGAG TCGTGATGTC AGGTTTTCGT GGCAGACGGC CAAAGATTGT
     2341 CTTGAGAGAC ACAACTCAAT CAAAGTTGTG TTTGGCGAAG AGGAGCCTAC CCCGGTTAAT
     2401 TTCGGTGATC TGGGGTTGAG CAAACAGAAG CCCGACTTTA AGGATTTAGA AATCAGTACC
     2461 AGTTGGTGGG GACATTAATT GGTGGAGGAG AGCCAATATT CTCAACTGTT CGTAGGACCA
     2521 TAACCCCCCA CTCGCACCAG AAATACTCAT TGGCGCTGAC AACTAGTTTG AGCCGCTTCA
     2581 ATTGCGGCAA GTCCCCAGTT TTGTTTGGTA CTCCCCTCTC GCCTCGCGTG TTTTTGTTCC
     2641 TCCGAATTGT GCATCACCCT GTCACATCCG TACACCATCG GAACACTCTG CTCCTCCTCC
     2701 TTTCTCCTCT TAATTGAACG TATATCTAAT TACCCTAATA ACAAGTATCT TCAACCATGG
     2761 ACAAGAAGTA CTCCATTGGG CTCGATATCG GCACAAACAG CGTCGGTTGG GCCGTCATTA
     2821 CGGACGAGTA CAAGGTGCCG AGCAAAAAAT TCAAAGTTCT GGGCAATACC GATCGCCACA
     2881 GCATAAAGAA GAACCTCATT GGCGCCCTCC TGTTCGACTC CGGGGAGACG GCCGAAGCCA
     2941 CGCGGCTCAA AAGAACAGCA CGGCGCAGAT ATACCCGCAG AAAGAATCGG ATCTGCTACC
     3001 TGCAGGAGAT CTTTAGTAAT GAGATGGCTA AGGTGGATGA CTCTTTCTTC CATAGGCTGG
     3061 AGGAGTCCTT TTTGGTGGAG GAGGATAAAA AGCACGAGCG CCACCCAATC TTTGGCAATA
     3121 TCGTGGACGA GGTGGCGTAC CATGAAAAGT ACCCAACCAT ATATCATCTG AGGAAGAAGC
     3181 TTGTAGACAG TACTGATAAG GCTGACTTGC GGTTGATCTA TCTCGCGCTG GCGCATATGA
     3241 TCAAATTTCG GGGACACTTC CTCATCGAGG GGGACCTGAA CCCAGACAAC AGCGATGTCG
     3301 ACAAACTCTT TATCCAACTG GTTCAGACTT ACAATCAGCT TTTCGAAGAG AACCCGATCA
     3361 ACGCATCCGG AGTTGACGCC AAAGCAATCC TGAGCGCTAG GCTGTCCAAA TCCCGGCGGC
     3421 TCGAAAACCT CATCGCACAG CTCCCTGGGG AGAAGAAGAA CGGCCTGTTT GGTAATCTTA
     3481 TCGCCCTGTC ACTCGGGCTG ACCCCCAACT TTAAATCTAA CTTCGACCTG GCCGAAGATG
     3541 CCAAGCTTCA ACTGAGCAAA GACACCTACG ATGATGATCT CGACAATCTG CTGGCCCAGA
     3601 TCGGCGACCA GTACGCAGAC CTTTTTTTGG CGGCAAAGAA CCTGTCAGAC GCCATTCTGC
     3661 TGAGTGATAT TCTGCGAGTG AACACGGAGA TCACCAAAGC TCCGCTGAGC GCTAGTATGA
     3721 TCAAGCGCTA TGATGAGCAC CACCAAGACT TGACTTTGCT GAAGGCCCTT GTCAGACAGC
     3781 AACTGCCTGA GAAGTACAAG GAAATTTTCT TCGATCAGTC TAAAAATGGC TACGCCGGAT
     3841 ACATTGACGG CGGAGCAAGC CAGGAGGAAT TTTACAAATT TATTAAGCCC ATCTTGGAAA
     3901 AAATGGACGG CACCGAGGAG CTGCTGGTAA AGCTTAACAG AGAAGATCTG TTGCGCAAAC
     3961 AGCGCACTTT CGACAATGGA AGCATCCCCC ACCAGATTCA CCTGGGCGAA CTGCACGCTA
     4021 TCCTCAGGCG GCAAGAGGAT TTCTACCCCT TTTTGAAAGA TAACAGGGAA AAGATTGAGA
     4081 AAATCCTCAC ATTTCGGATA CCCTACTATG TAGGCCCCCT CGCCCGGGGA AATTCCAGAT
     4141 TCGCGTGGAT GACTCGCAAA TCAGAAGAGA CtATCACTCC CTGGAACTTC GAGGAAGTCG
     4201 TGGATAAGGG GGCCTCTGCC CAGTCCTTCA TCGAAAGGAT GACTAACTTT GATAAAAATC
     4261 TGCCTAACGA AAAGGTGCTT CCTAAACACT CTCTGCTGTA CGAGTACTTC ACAGTTTATA
     4321 ACGAGCTCAC CAAGGTCAAA TACGTCACAG AAGGGATGAG AAAGCCAGCA TTCCTGTCTG
     4381 GAGAGCAGAA GAAAGCTATC GTGGACCTCC TCTTCAAGAC GAACCGGAAA GTTACCGTGA
     4441 AACAGCTCAA AGAgGACTAT TTCAAAAAGA TTGAATGTTT CGACTCTGTT GAAATCAGCG
     4501 GAGTGGAGGA TCGCTTCAAC GCATCCCTGG GAACGTATCA CGATCTCCTG AAAATCATTA
     4561 AAGACAAGGA CTTCCTGGAC AATGAGGAGA ACGAGGACAT TCTTGAGGAC ATTGTCCTCA
     4621 CCCTTACGTT GTTTGAAGAT AGGGAGATGA TTGAAGAACG CTTGAAAACT TACGCTCATC
     4681 TCTTCGACGA CAAAGTCATG AAACAGCTCA AGAGGCGCCG ATATACAGGA TGGGGGCGGC
     4741 TGTCAAGAAA ACTGATCAAT GGGATCCGAG ACAAGCAGAG TGGAAAGACA ATCCTGGATT
     4801 TTCTTAAGTC CGATGGATTT GCCAACCGGA ACTTCATGCA GTTGATCCAT GATGACTCTC
     4861 TCACCTTTAA GGAGGACATC CAGAAAGCAC AAGTTTCTGG CCAGGGGGAC AGcCTTCACG
     4921 AGCACATCGC TAATCTTGCA GGTAGCCCAG CTATCAAAAA GGGAATACTG CAGACCGTTA
     4981 AGGTCGTGGA TGAACTCGTC AAAGTAATGG GAAGGCATAA GCCCGAGAAT ATCGTTATCG
     5041 AGATGGCCCG AGAGAACCAA ACTACCCAGA AGGGACAGAA GAACAGTAGG GAAAGGATGA
     5101 AGAGGATTGA AGAGGGTATA AAAGAACTGG GGTCCCAAAT CCTTAAGGAA CACCCAGTTG
     5161 AAAACACCCA GCTTCAGAAT GAGAAGCTCT ACCTGTACTA CCTGCAGAAC GGCAGGGACA
     5221 TGTACGTGGA TCAGGAACTG GACATCAATC GGCTCTCCGA CTACGACGTG GATCATATCG
     5281 TGCCCCAGTC TTTTCTCAAA GATGATTCTA TTGATAATAA AGTGTTGACA AGATCCGATA
     5341 AAAATAGAGG GAAGAGTGAT AACGTCCCCT CAGAAGAAGT TGTCAAGAAA ATGAAAAATT
     5401 ATTGGCGGCA GCTGCTGAAC GCCAAACTGA TCACACAACG GAAGTTCGAT AATCTGACTA
     5461 AGGCTGAACG AGGTGGCCTG TCTGAGTTGG ATAAAGCCGG CTTCATCAAA AGGCAGCTTG
     5521 TTGAGACACG CCAGATCACC AAGCACGTGG CCCAAATTCT CGATTCACGC ATGAACACCA
     5581 AGTACGATGA AAATGACAAA CTGATTCGAG AGGTGAAAGT TATTACTCTG AAGTCTAAGC
     5641 TcGTCTCAGA TTTCAGAAAG GACTTTCAGT TTTATAAGGT GAGAGAGATC AACAATTACC
     5701 ACCATGCGCA TGATGCCTAC CTGAATGCAG TGGTAGGCAC TGCACTTATC AAAAAATATC
     5761 CCAAGCTTGA ATCTGAATTT GTTTACGGAG ACTATAAAGT GTACGATGTT AGGAAAATGA
     5821 TCGCAAAGTC TGAGCAGGAA ATAGGCAAGG CCACCGCTAA GTACTTCTTT TACAGCAATA
     5881 TTATGAATTT TTTCAAGACC GAGATTACAC TGGCCAATGG AGAGATTCGG AAGCGACCAC
     5941 TTATCGAAAC AAACGGAGAA ACAGGAGAAA TCGTGTGGGA CAAGGGTAGG GATTTCGCGA
     6001 CAGTCCGGAA GGTCCTGTCC ATGCCGCAGG TGAACATCGT TAAAAAGACC GAAGTACAGA
     6061 CCGGAGGCTT CTCCAAGGAA AGTATCCTCC CGAAAAGGAA CAGCGACAAG CTGATCGCAC
     6121 GCAAAAAAGA TTGGGACCCC AAGAAATACG GCGGATTCGA TTCTCCTACA GTCGCTTACA
     6181 GTGTACTGGT TGTGGCCAAA GTGGAGAAAG GGAAGTCTAA AAAACTCAAA AGCGTCAAGG
     6241 AACTGCTGGG CATCACAATC ATGGAGCGAT CAAGCTTCGA AAAAAACCCC ATCGACTTTC
     6301 TCGAGGCGAA AGGATATAAA GAGGTCAAAA AAGACCTCAT CATTAAGCTT CCCAAGTACT
     6361 CTCTCTTTGA GCTTGAAAAC GGCCGGAAAC GAATGCTCGC TAGTGCGGGC GAGCTGCAGA
     6421 AAGGTAACGA GCTGGCACTG CCCTCTAAAT ACGTTAATTT CTTGTATCTG GCCAGCCACT
     6481 ATGAAAAGCT CAAAGGGTCc CCCGAAGATA ATGAGCAGAA GCAGCTGTTC GTGGAACAAC
     6541 ACAAACACTA CCTTGATGAG ATCATCGAGC AAATAAGCGA ATTCTCCAAA AGAGTGATCC
     6601 TCGCCGACGC TAACCTCGAT AAGGTGCTTT CTGCTTACAA TAAGCACAGG GATAAGCCCA
     6661 TCAGGGAGCA GGCAGAAAAC ATTATCCACT TGTTTACTCT GACCAACTTG GGCGCGCCTG
     6721 CAGCCTTCAA GTACTTCGAC ACCACCATAG ACAGAAAGCG GTACACCTCT ACAAAGGAGG
     6781 TCCTGGACGC CACACTGATT CATCAGTCAA TTACGGGGCT CTATGAAACA AGAATCGACC
     6841 TCTCTCAGCT CGGTGGAGAC AGCAGGGCTG ACCCCAAGAA GAAGAGGAAG GTGTGAGCTT
     6901 CGTCCGACGG CGGCCCACGG GTCCCAGGCC TCGGAGATCC GTCCCCCTTT TCCTTTGTCG
     6961 ATATCATGTA ATTAGTTATG TCACGCTTAC ATTCACGCCC TCCCCCCACA TCCGCTCTAA
     7021 CCGAAAAGGA AGGAGTTAGA CAACCTGAAG TCTAGGTCCC TATTTATTTT TTTATAGTTA
     7081 TGTTAGTATT AAGAACGTTA TTTATATTTC AAATTTTTCT TTTTTTTCTG TACAGACGCG
     7141 TGTACGCATG TAACATTATA CTGAAAACCT TGCTTGAGAA GGTTTTGGGA CGCTCGAAGG
     7201 CTTTAATTTG CAAGCTGGCG CTAATTCATG GACATGGAGG CCCAGAATAC CCTCCTTGAC
     7261 AGTCTTGACG TGCGCAGCTC AGGGGCATGA TGTGACTGTC GCCCGTACAT TTAGCCCATA
     7321 CATCCCCATG TATAATCATT TGCATCCATA CATTTTGATG GCCGCACGGC GCGAAGCAAA
     7381 AATTACGGCT CCTCGCTGCA GACCTGCGAG CAGGGAAACG CTCCCCTCAC AGACGCGTTG
     7441 AATTGTCCCC ACGCCGCGCC CCTGTAGAGA AATATAAAAG GTTAGGATTT GCCACTGAGG
     7501 TTCTTCTTTC ATATACTTCC TTTTAAAATC TTGCTAGGAT ACAGTTCTCA CATCACATCC
     7561 GAACATAAAC AACCATGggt accactcttg acgacacggc ttaccggtac cgcaccagtg
     7621 tcccggggga cgccgaggcc atcgaggcac tggatgggtc cttcaccacc gacaccgtTT
     7681 TTcgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg gacccgcccc
     7741 tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg gaggacggcg
     7801 acccggactc ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg ggcttcgtgg
     7861 tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag gtcgccccgg
     7921 agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag ttcgcccgcg
     7981 agcggggcgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg gcgatccacg
     8041 cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac gacggcaccg
     8101 cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaatca gtACTGACAA
     8161 TAAAAAGATT CTTGTTTTCA AGAACTTGTC ATTTGTATAG TTTTTTTATA TTGTAGTTGT
     8221 TCTATTTTAA TCAAATGTTA GCGTGATTTA TATTTTTTTT CGCCTCGACA TCATCTGCCC
     8281 AGATGCGAAG TTAAGTGCGC AGAAAGTAAT ATCATGCGTC AATCGTATGT GAATGCTGGT
     8341 CGCTATACTG GCTTCCTTTT CATCACGTGC TATAAAAATA ATTATAATTT AAATTTTTTA
     8401 ATATAAATAT ATAAATTAAA AATAGAAAGT AAAAAAAGAA ATTAAAGAAA AAATAGTTTT
     8461 TGTTTTCCGA AGATGTAAAA GACTCTAGGG GGATCGCCAA CAAATACTAC CTTTTATCTT
     8521 GCTCTTCCTG CTCTCAGGTA TTAATGCCGA ATTGTTTCAT CTTGTCTGTG TAGAACACCA
     8581 CACACGAAAA TCCTGTGATT TTACATTTTA CTTATCGTTA ATCGAATGTA TATCTATTTA
     8641 ATCTGCTTTT CTTGTCTAAT AAATATATAT GTAAAGTACG CTTTTTGTTG AAATTTTTTA
     8701 AACCTTTGTT TATTTTTTTT TCTTCATTCC GTAACTCTTC TACCTTCTTT ATTTACTTTC
     8761 TAAAATCCAA ATACAAAACA TAAAAATAAA TAAACACAGA GTAAATTCCC AAATTATTCC
     8821 ATCATTAAAA GATACGAGGC GCGTGTAAGT TACAGGCAAG CGATCG
//
LOCUS       BB3_pGAP_Linker_BbsI(intern)_RPS25Att_pPFK300_Cas9_cycTT_Hyg      8545 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Vector          join(6446..8545,1..722)
                     /origin="183_BB3_Hygro_FsA_FsC_CEN6-ARS4 (2,852bp
                     circular)Target vector: from BsaI cut at 752^753 to BsaI
                     cut at 722^723, 2,822bpOrientation: originalPlus strand
                     5':   no changePlus strand 3':   no changeMinus strand
                     5':  no changeMinus strand 3':  no change"
     rep_origin      complement(5..678)
                     /label="pUC ORI"
     terminator      683..722
                     /label="BBa_B1007"
     FS              723..726
                     /label="FS A"
     Vector          join(1743..>1746,<723..726)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 726^727
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        729..>1221
                     /label="pGAP"
     Restriction_sit join(1221^1222,complement(1221^1222))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1222..>1253
                     /label="L_FS23_BbsI_fw"
     Linker          1222..1253
                     /origin="PCR_Linker_FS23_BbsI(intern) (54bp
                     linear)Fragment: from BpiI cut at 9^10 to BpiI cut at
                     41^42, 32bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="FS 23 Linker"
     FS              1222..1225
                     /label="FS2"
     primer          complement(<1226..>1257)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1254..1286,complement(1287..1719),complement(1720..>1746))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1258..>1742
                     /note="Forward primer name: RPS25ATT_M1_FW"
                     /note="Forward primer: GacagGtcTccgCTTCCTTGTCGAGTGTGTTT"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_RV"
                     /note="Reverse primer:
                     gatcggtctCcAgctGTCAAATATTCAGATGGTGA"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 458"
                     /label="rps25aTT"
     FS              1743..1746
                     /label="FS 4"
     primer          <1747..>1750
                     /label="3_BsaI_FsB_Bam"
     misc_feature    join(<6442..>6445,<1747..>1750)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (108bp linear)Fragment: full
                     sequenceOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1747..1750
                     /label="FS B"
     Vector          join(6442..>6445,<1747..1750)
                     /origin="BB2_L_BC_syn_BbsI_7 (2,215bp circular)Target
                     vector: from BpiI cut at 639^640 to BpiI cut at 607^608,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          complement(<1751..>1754)
                     /label="4_BsaI_FsB_BamXho"
     promoter        1763..1978
                     /label="pPFK300"
     primer          <1979..2003
                     /label="FS2_Cas9_BsaI_fw"
     CDS             1980..6119
                     /label="Cas9"
     Taming          2030
                     /label="C>T"
     primer          complement(3391..3410)
                     /label="1_cas9_rev"
     primer          3391..3410
                     /label="1_cas9_fw"
     Taming          3395
                     /label="C>t"
     primer          complement(3665..3686)
                     /label="2_cas9_rev"
     primer          3665..3686
                     /label="2_cas9_fw"
     Taming          3677
                     /label="A>g"
     primer          complement(4134..4151)
                     /label="3_cas9_rev"
     primer          4134..4151
                     /label="3_cas9_fw"
     Taming          4136
                     /label="T>c"
     primer          complement(4849..4870)
                     /label="4_cas9_rev"
     primer          4849..4870
                     /label="4_cas9_fw"
     Taming          4865
                     /label="G>c"
     primer          complement(5716..5733)
                     /label="5_cas9_rev"
     primer          5716..5734
                     /label="5_cas9_fw"
     Taming          5723
                     /label="T>c"
     NLS             6084..6119
                     /label="NLS SV40"
     primer          complement(6097..>6123)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6120..>6441
                     /label="FS3_CYCtt_FS4"
     primer          <6442..>6445
                     /label="5_BsaI_FsC_Xho"
     FS              6442..6445
                     /label="FS 4"
     primer          complement(6446..>6449)
                     /label="6_BsaI_FsC"
     promoter        6454..6797
                     /label="TEFpromoter"
     CDS             6798..7826
                     /label="hphR (Hygromycin)"
     terminator      7835..8029
                     /label="TEFterminator"
     misc_feature    8034..8545
                     /label="CEN6_ARS4"
     mutation        8255
                     /label="G/A"
ORIGIN
        1 CGCTGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT
       61 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT
      121 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC
      181 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA
      241 GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT
      301 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA
      361 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG
      421 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC
      481 CTAACTACGG CTACACTAGA AGAACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA
      541 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG
      601 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT
      661 TGATCTTTTC TACGGGGTGG AGcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcGATCGGAG GATCCTTTTT TGTAGAAATG TCTTGGTGTC CTCGTCCAAT CAGGTAGCCA
      781 TCTCTGAAAT ATCTGGCTCC GTTGCAACTC CGAACGACCT GCTGGCAACG TAAAATTCTC
      841 CGGGGTAAAA CTTAAATGTG GAGTAATGGA ACCAGAAACG TCTCTTCCCT TCTCTCTCCT
      901 TCCACCGCCC GTTACCGTCC CTAGGAAATT TTACTCTGCT GGAGAGCTTC TTCTACGGCC
      961 CCCTTGCAGC AATGCTCTTC CCAGCATTAC GTTGCGGGTA AAACGGAGGT CGTGTACCCG
     1021 ACCTAGCAGC CCAGGGATGG AAAAGTCCCG GCCGTCGCTG GCAATAATAG CGGGCGGACG
     1081 CATGTCATGA GATTATTGGA AACCACCAGA ATCGAATATA AAAGGCGAAC ACCTTTCCCA
     1141 ATTTTGGTTT CTCCTGACCC AAAGACTTTA AATTTAATTT ATTTGTCCCT ATTTCAATCA
     1201 ATTGAACAAC TATCAAAACA CCATGGCGTC TTCGGATCCC TCGAGGAAGA CGCGCTTTTA
     1261 GTGTACATCT GATAATATAG TACTACCACG TATGATAATG TAGAGAATAG GCTTCCTTGT
     1321 CGAGTGTGTT TGCAGTTTTC TTGAGTTTCA AGGTTTAAAT GCTGGTATAT TAGTTCATCG
     1381 AAGGTTTCAG CCAATAGCAC CTTAAATCAA TCAAACTAAT TCGACTCTTA CGAAAGAGCC
     1441 TACTGTGTTT AGTATCGAAG TCGTTTACCT TTCATGTTGA ATAGCTTCCT CTCTGACCCT
     1501 AACATTTCAA GATCCTCCTA AAGTTACCCG GATTGTGAAA TTCTAATGAT CCACCTGCCC
     1561 AATGCATTTT TTCTTTATTC AGTTTACCTT TTTTACCTAA TATACGAGCT TGTTAAAGTA
     1621 AGTGGCACTG CAATACTAGG CTTATTGTTG ATATTATGAT GAATCGTTTT CACAAACTTG
     1681 ATTTCCTGTG AACTCACCAT GTACTAAGGA AAAAAACATG CATCACCATC TGAATATTTG
     1741 ACCGCTCCGG GGAGCCTGCC ATTAGGCAAA CACCAACTAG GAACCTACAC CAAGTTATGA
     1801 AGGAGCTTGG AAAGGATTAT AAAAAATGCG ATTACCTAAT ACACACACGA TGCCTGAGCG
     1861 GTTAATGTGA AATTTCCCGG CCATCGATCG CCCCACAATC AGACTATTTT CGCTCATACA
     1921 ATAAAAAGTC TCCTGTTTCC ACCCGAGTAT CACCAGAACA AAGTAACAAA TACAAAACCA
     1981 TGGACAAGAA GTACTCCATT GGGCTCGATA TCGGCACAAA CAGCGTCGGT TGGGCCGTCA
     2041 TTACGGACGA GTACAAGGTG CCGAGCAAAA AATTCAAAGT TCTGGGCAAT ACCGATCGCC
     2101 ACAGCATAAA GAAGAACCTC ATTGGCGCCC TCCTGTTCGA CTCCGGGGAG ACGGCCGAAG
     2161 CCACGCGGCT CAAAAGAACA GCACGGCGCA GATATACCCG CAGAAAGAAT CGGATCTGCT
     2221 ACCTGCAGGA GATCTTTAGT AATGAGATGG CTAAGGTGGA TGACTCTTTC TTCCATAGGC
     2281 TGGAGGAGTC CTTTTTGGTG GAGGAGGATA AAAAGCACGA GCGCCACCCA ATCTTTGGCA
     2341 ATATCGTGGA CGAGGTGGCG TACCATGAAA AGTACCCAAC CATATATCAT CTGAGGAAGA
     2401 AGCTTGTAGA CAGTACTGAT AAGGCTGACT TGCGGTTGAT CTATCTCGCG CTGGCGCATA
     2461 TGATCAAATT TCGGGGACAC TTCCTCATCG AGGGGGACCT GAACCCAGAC AACAGCGATG
     2521 TCGACAAACT CTTTATCCAA CTGGTTCAGA CTTACAATCA GCTTTTCGAA GAGAACCCGA
     2581 TCAACGCATC CGGAGTTGAC GCCAAAGCAA TCCTGAGCGC TAGGCTGTCC AAATCCCGGC
     2641 GGCTCGAAAA CCTCATCGCA CAGCTCCCTG GGGAGAAGAA GAACGGCCTG TTTGGTAATC
     2701 TTATCGCCCT GTCACTCGGG CTGACCCCCA ACTTTAAATC TAACTTCGAC CTGGCCGAAG
     2761 ATGCCAAGCT TCAACTGAGC AAAGACACCT ACGATGATGA TCTCGACAAT CTGCTGGCCC
     2821 AGATCGGCGA CCAGTACGCA GACCTTTTTT TGGCGGCAAA GAACCTGTCA GACGCCATTC
     2881 TGCTGAGTGA TATTCTGCGA GTGAACACGG AGATCACCAA AGCTCCGCTG AGCGCTAGTA
     2941 TGATCAAGCG CTATGATGAG CACCACCAAG ACTTGACTTT GCTGAAGGCC CTTGTCAGAC
     3001 AGCAACTGCC TGAGAAGTAC AAGGAAATTT TCTTCGATCA GTCTAAAAAT GGCTACGCCG
     3061 GATACATTGA CGGCGGAGCA AGCCAGGAGG AATTTTACAA ATTTATTAAG CCCATCTTGG
     3121 AAAAAATGGA CGGCACCGAG GAGCTGCTGG TAAAGCTTAA CAGAGAAGAT CTGTTGCGCA
     3181 AACAGCGCAC TTTCGACAAT GGAAGCATCC CCCACCAGAT TCACCTGGGC GAACTGCACG
     3241 CTATCCTCAG GCGGCAAGAG GATTTCTACC CCTTTTTGAA AGATAACAGG GAAAAGATTG
     3301 AGAAAATCCT CACATTTCGG ATACCCTACT ATGTAGGCCC CCTCGCCCGG GGAAATTCCA
     3361 GATTCGCGTG GATGACTCGC AAATCAGAAG AGACtATCAC TCCCTGGAAC TTCGAGGAAG
     3421 TCGTGGATAA GGGGGCCTCT GCCCAGTCCT TCATCGAAAG GATGACTAAC TTTGATAAAA
     3481 ATCTGCCTAA CGAAAAGGTG CTTCCTAAAC ACTCTCTGCT GTACGAGTAC TTCACAGTTT
     3541 ATAACGAGCT CACCAAGGTC AAATACGTCA CAGAAGGGAT GAGAAAGCCA GCATTCCTGT
     3601 CTGGAGAGCA GAAGAAAGCT ATCGTGGACC TCCTCTTCAA GACGAACCGG AAAGTTACCG
     3661 TGAAACAGCT CAAAGAgGAC TATTTCAAAA AGATTGAATG TTTCGACTCT GTTGAAATCA
     3721 GCGGAGTGGA GGATCGCTTC AACGCATCCC TGGGAACGTA TCACGATCTC CTGAAAATCA
     3781 TTAAAGACAA GGACTTCCTG GACAATGAGG AGAACGAGGA CATTCTTGAG GACATTGTCC
     3841 TCACCCTTAC GTTGTTTGAA GATAGGGAGA TGATTGAAGA ACGCTTGAAA ACTTACGCTC
     3901 ATCTCTTCGA CGACAAAGTC ATGAAACAGC TCAAGAGGCG CCGATATACA GGATGGGGGC
     3961 GGCTGTCAAG AAAACTGATC AATGGGATCC GAGACAAGCA GAGTGGAAAG ACAATCCTGG
     4021 ATTTTCTTAA GTCCGATGGA TTTGCCAACC GGAACTTCAT GCAGTTGATC CATGATGACT
     4081 CTCTCACCTT TAAGGAGGAC ATCCAGAAAG CACAAGTTTC TGGCCAGGGG GACAGcCTTC
     4141 ACGAGCACAT CGCTAATCTT GCAGGTAGCC CAGCTATCAA AAAGGGAATA CTGCAGACCG
     4201 TTAAGGTCGT GGATGAACTC GTCAAAGTAA TGGGAAGGCA TAAGCCCGAG AATATCGTTA
     4261 TCGAGATGGC CCGAGAGAAC CAAACTACCC AGAAGGGACA GAAGAACAGT AGGGAAAGGA
     4321 TGAAGAGGAT TGAAGAGGGT ATAAAAGAAC TGGGGTCCCA AATCCTTAAG GAACACCCAG
     4381 TTGAAAACAC CCAGCTTCAG AATGAGAAGC TCTACCTGTA CTACCTGCAG AACGGCAGGG
     4441 ACATGTACGT GGATCAGGAA CTGGACATCA ATCGGCTCTC CGACTACGAC GTGGATCATA
     4501 TCGTGCCCCA GTCTTTTCTC AAAGATGATT CTATTGATAA TAAAGTGTTG ACAAGATCCG
     4561 ATAAAAATAG AGGGAAGAGT GATAACGTCC CCTCAGAAGA AGTTGTCAAG AAAATGAAAA
     4621 ATTATTGGCG GCAGCTGCTG AACGCCAAAC TGATCACACA ACGGAAGTTC GATAATCTGA
     4681 CTAAGGCTGA ACGAGGTGGC CTGTCTGAGT TGGATAAAGC CGGCTTCATC AAAAGGCAGC
     4741 TTGTTGAGAC ACGCCAGATC ACCAAGCACG TGGCCCAAAT TCTCGATTCA CGCATGAACA
     4801 CCAAGTACGA TGAAAATGAC AAACTGATTC GAGAGGTGAA AGTTATTACT CTGAAGTCTA
     4861 AGCTcGTCTC AGATTTCAGA AAGGACTTTC AGTTTTATAA GGTGAGAGAG ATCAACAATT
     4921 ACCACCATGC GCATGATGCC TACCTGAATG CAGTGGTAGG CACTGCACTT ATCAAAAAAT
     4981 ATCCCAAGCT TGAATCTGAA TTTGTTTACG GAGACTATAA AGTGTACGAT GTTAGGAAAA
     5041 TGATCGCAAA GTCTGAGCAG GAAATAGGCA AGGCCACCGC TAAGTACTTC TTTTACAGCA
     5101 ATATTATGAA TTTTTTCAAG ACCGAGATTA CACTGGCCAA TGGAGAGATT CGGAAGCGAC
     5161 CACTTATCGA AACAAACGGA GAAACAGGAG AAATCGTGTG GGACAAGGGT AGGGATTTCG
     5221 CGACAGTCCG GAAGGTCCTG TCCATGCCGC AGGTGAACAT CGTTAAAAAG ACCGAAGTAC
     5281 AGACCGGAGG CTTCTCCAAG GAAAGTATCC TCCCGAAAAG GAACAGCGAC AAGCTGATCG
     5341 CACGCAAAAA AGATTGGGAC CCCAAGAAAT ACGGCGGATT CGATTCTCCT ACAGTCGCTT
     5401 ACAGTGTACT GGTTGTGGCC AAAGTGGAGA AAGGGAAGTC TAAAAAACTC AAAAGCGTCA
     5461 AGGAACTGCT GGGCATCACA ATCATGGAGC GATCAAGCTT CGAAAAAAAC CCCATCGACT
     5521 TTCTCGAGGC GAAAGGATAT AAAGAGGTCA AAAAAGACCT CATCATTAAG CTTCCCAAGT
     5581 ACTCTCTCTT TGAGCTTGAA AACGGCCGGA AACGAATGCT CGCTAGTGCG GGCGAGCTGC
     5641 AGAAAGGTAA CGAGCTGGCA CTGCCCTCTA AATACGTTAA TTTCTTGTAT CTGGCCAGCC
     5701 ACTATGAAAA GCTCAAAGGG TCcCCCGAAG ATAATGAGCA GAAGCAGCTG TTCGTGGAAC
     5761 AACACAAACA CTACCTTGAT GAGATCATCG AGCAAATAAG CGAATTCTCC AAAAGAGTGA
     5821 TCCTCGCCGA CGCTAACCTC GATAAGGTGC TTTCTGCTTA CAATAAGCAC AGGGATAAGC
     5881 CCATCAGGGA GCAGGCAGAA AACATTATCC ACTTGTTTAC TCTGACCAAC TTGGGCGCGC
     5941 CTGCAGCCTT CAAGTACTTC GACACCACCA TAGACAGAAA GCGGTACACC TCTACAAAGG
     6001 AGGTCCTGGA CGCCACACTG ATTCATCAGT CAATTACGGG GCTCTATGAA ACAAGAATCG
     6061 ACCTCTCTCA GCTCGGTGGA GACAGCAGGG CTGACCCCAA GAAGAAGAGG AAGGTGTGAG
     6121 CTTCGTCCGA CGGCGGCCCA CGGGTCCCAG GCCTCGGAGA TCCGTCCCCC TTTTCCTTTG
     6181 TCGATATCAT GTAATTAGTT ATGTCACGCT TACATTCACG CCCTCCCCCC ACATCCGCTC
     6241 TAACCGAAAA GGAAGGAGTT AGACAACCTG AAGTCTAGGT CCCTATTTAT TTTTTTATAG
     6301 TTATGTTAGT ATTAAGAACG TTATTTATAT TTCAAATTTT TCTTTTTTTT CTGTACAGAC
     6361 GCGTGTACGC ATGTAACATT ATACTGAAAA CCTTGCTTGA GAAGGTTTTG GGACGCTCGA
     6421 AGGCTTTAAT TTGCAAGCTG GCGCTAATTC ATGGACATGG AGGCCCAGAA TACCCTCCTT
     6481 GACAGTCTTG ACGTGCGCAG CTCAGGGGCA TGATGTGACT GTCGCCCGTA CATTTAGCCC
     6541 ATACATCCCC ATGTATAATC ATTTGCATCC ATACATTTTG ATGGCCGCAC GGCGCGAAGC
     6601 AAAAATTACG GCTCCTCGCT GCAGACGTGC GAGCAGGGAA ACGCTCCCCT CACAGACGCG
     6661 TTGAATTGTC CCCACGCCGC GCCCCTGTAG AGAAATATAA AAGGTTAGGA TTTGCCACTG
     6721 AGGTTCTTCT TTCATATACT TCCTTTTAAA ATCTTGCTAG GATACAGTTC TCACATCACA
     6781 TCCGAACATA AACAACCATG GGTAAAAAGC CTGAACTCAC CGCGACGTCT GTCGAGAAGT
     6841 TTCTGATCGA AAAGTTCGAC AGCGTCTCCG ACCTGATGCA GCTCTCGGAG GGCGAAGAAT
     6901 CTCGTGCTTT CAGCTTCGAT GTAGGAGGGC GTGGATATGT CCTGCGGGTA AATAGCTGCG
     6961 CCGATGGTTT CTACAAAGAT CGTTATGTTT ATCGGCACTT TGCATCGGCC GCGCTCCCGA
     7021 TTCCGGAAGT GCTTGACATT GGGGAATTCA GCGAGAGCCT GACCTATTGC ATCTCCCGCC
     7081 GTGCACAGGG TGTCACGTTG CAAGACCTCC CTGAAACCGA ACTGCCCGCT GTTCTGCAGC
     7141 CGGTCGCGGA GGCCATGGAT GCGATCGCTG CGGCCGATCT TAGCCAGACG AGCGGGTTCG
     7201 GCCCATTCGG ACCGCAAGGA ATCGGTCAAT ACACTACATG GCGTGATTTC ATATGCGCGA
     7261 TTGCTGATCC CCATGTGTAT CACTGGCAAA CTGTGATGGA CGACACCGTC AGTGCGTCCG
     7321 TCGCGCAGGC TCTCGATGAG CTGATGCTTT GGGCCGAGGA CTGCCCCGAA GTCCGGCACC
     7381 TCGTGCACGC GGATTTCGGC TCCAACAATG TCCTGACGGA CAATGGCCGC ATAACAGCGG
     7441 TCATTGACTG GAGCGAGGCG ATGTTCGGGG ATTCCCAATA CGAGGTCGCC AACATCTTCT
     7501 TCTGGAGGCC GTGGTTGGCT TGTATGGAGC AGCAGACGCG CTACTTCGAG CGGAGGCATC
     7561 CGGAGCTTGC AGGATCGCCG CGGCTCCGGG CGTATATGCT CCGCATTGGT CTTGACCAAC
     7621 TCTATCAGAG CTTGGTTGAC GGCAATTTCG ATGATGCAGC TTGGGCGCAG GGTCGATGCG
     7681 ACGCAATCGT CCGATCCGGA GCCGGGACTG TCGGGCGTAC ACAAATCGCC CGCAGAAGCG
     7741 CGGCCGTCTG GACCGATGGC TGTGTAGAAG TACTCGCCGA TAGTGGAAAC CGACGCCCCA
     7801 GCACTCGTCC GAGGGCAAAG GAATAATCAG TACTGACAAT AAAAAGATTC TTGTTTTCAA
     7861 GAACTTGTCA TTTGTATAGT TTTTTTATAT TGTAGTTGTT CTATTTTAAT CAAATGTTAG
     7921 CGTGATTTAT ATTTTTTTTC GCCTCGACAT CATCTGCCCA GATGCGAAGT TAAGTGCGCA
     7981 GAAAGTAATA TCATGCGTCA ATCGTATGTG AATGCTGGTC GCTATACTGG CTTCCTTTTC
     8041 ATCACGTGCT ATAAAAATAA TTATAATTTA AATTTTTTAA TATAAATATA TAAATTAAAA
     8101 ATAGAAAGTA AAAAAAGAAA TTAAAGAAAA AATAGTTTTT GTTTTCCGAA GATGTAAAAG
     8161 ACTCTAGGGG GATCGCCAAC AAATACTACC TTTTATCTTG CTCTTCCTGC TCTCAGGTAT
     8221 TAATGCCGAA TTGTTTCATC TTGTCTGTGT AGAACACCAC ACACGAAAAT CCTGTGATTT
     8281 TACATTTTAC TTATCGTTAA TCGAATGTAT ATCTATTTAA TCTGCTTTTC TTGTCTAATA
     8341 AATATATATG TAAAGTACGC TTTTTGTTGA AATTTTTTAA ACCTTTGTTT ATTTTTTTTT
     8401 CTTCATTCCG TAACTCTTCT ACCTTCTTTA TTTACTTTCT AAAATCCAAA TACAAAACAT
     8461 AAAAATAAAT AAACACAGAG TAAATTCCCA AATTATTCCA TCATTAAAAG ATACGAGGCG
     8521 CGTGTAAGTT ACAGGCAAGC GATCG
//
LOCUS       BB3_pGAP_Linker_BbsI(intern)_RPS25Att_pLAT1_Cas9_cycTT_Hyg      9322 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Vector          join(7223..9322,1..722)
                     /origin="183_BB3_Hygro_FsA_FsC_CEN6-ARS4 (2,852bp
                     circular)Target vector: from BsaI cut at 752^753 to BsaI
                     cut at 722^723, 2,822bpOrientation: originalPlus strand
                     5':   no changePlus strand 3':   no changeMinus strand
                     5':  no changeMinus strand 3':  no change"
     rep_origin      complement(5..678)
                     /label="pUC ORI"
     terminator      683..722
                     /label="BBa_B1007"
     FS              723..726
                     /label="FS A"
     Vector          join(1743..>1746,<723..726)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 726^727
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        729..>1221
                     /label="pGAP"
     Restriction_sit join(1221^1222,complement(1221^1222))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1222..>1253
                     /label="L_FS23_BbsI_fw"
     Linker          1222..1253
                     /origin="PCR_Linker_FS23_BbsI(intern) (54bp
                     linear)Fragment: from BpiI cut at 9^10 to BpiI cut at
                     41^42, 32bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="FS 23 Linker"
     FS              1222..1225
                     /label="FS2"
     primer          complement(<1226..>1257)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1254..1286,complement(1287..1719),complement(1720..>1746))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1258..>1742
                     /note="Forward primer name: RPS25ATT_M1_FW"
                     /note="Forward primer: GacagGtcTccgCTTCCTTGTCGAGTGTGTTT"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_RV"
                     /note="Reverse primer:
                     gatcggtctCcAgctGTCAAATATTCAGATGGTGA"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 458"
                     /label="rps25aTT"
     FS              1743..1746
                     /label="FS 4"
     primer          <1747..>1750
                     /label="3_BsaI_FsB_Bam"
     misc_feature    join(<7219..>7222,<1747..>1750)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (108bp linear)Fragment: full
                     sequenceOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1747..1750
                     /label="FS B"
     Vector          join(7219..>7222,<1747..1750)
                     /origin="BB2_L_BC_syn_BbsI_7 (2,215bp circular)Target
                     vector: from BpiI cut at 639^640 to BpiI cut at 607^608,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          complement(<1751..>1754)
                     /label="4_BsaI_FsB_BamXho"
     promoter        <1751..>2755
                     /locus_tag="PAS_chr1-1_0049"
                     /codon_start=1
                     /product="Ribonuclease H2 catalytic subunit, removes RNA
                     primers during Okazaki fragment synthesis"
                     /protein_id="XP_002489688.1"
                     /db_xref="GI:254565155"
                     /db_xref="GeneID:8197566"
                     /translation="MTRIENRVCDSSPMTTTFIPPSVPEISTYRTTTYVSEIPESVLK
                     NPEKPCCLGVDEAGRGPVVGPMVYGVSYCLEEYEPRLKDFGFADSKTLTEQKRSELLE
                     SLCNGDHELRKNVGWATTSISACDISSAMLKPHNIGNYNLNEQAHDVTMELIDKVIQK
                     GVNLRHVYVDTVGPPASYQKKLQQRFPALLITVSKKADAIYPCVSSASVCAKVTRDVC
                     LRTSNGEENLLDCGSGYPSDPNTKKWLAQSIHPLFGWSRDVRFSWQTAKDCLERHNSI
                     KVVFGEEEPTPVNFGDLGLSKQKTDFKDLEISTSWWGH"
                     /label="pLAT1"
     Site            join(2755^2756,complement(2755^2756),complement(2755^2756))
                     /site_type="restriction site"
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 3162^3163"
                     /note="cut_0_on_negative_strand: 3166^3167"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <2756..2780
                     /label="FS2_Cas9_BsaI_fw"
     CDS             2757..6896
                     /label="Cas9"
     Taming          2807
                     /label="C>T"
     primer          complement(4168..4187)
                     /label="1_cas9_rev"
     primer          4168..4187
                     /label="1_cas9_fw"
     Taming          4172
                     /label="C>t"
     primer          complement(4442..4463)
                     /label="2_cas9_rev"
     primer          4442..4463
                     /label="2_cas9_fw"
     Taming          4454
                     /label="A>g"
     primer          complement(4911..4928)
                     /label="3_cas9_rev"
     primer          4911..4928
                     /label="3_cas9_fw"
     Taming          4913
                     /label="T>c"
     primer          complement(5626..5647)
                     /label="4_cas9_rev"
     primer          5626..5647
                     /label="4_cas9_fw"
     Taming          5642
                     /label="G>c"
     primer          complement(6493..6510)
                     /label="5_cas9_rev"
     primer          6493..6511
                     /label="5_cas9_fw"
     Taming          6500
                     /label="T>c"
     NLS             6861..6896
                     /label="NLS SV40"
     primer          complement(6874..>6900)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6897..>7218
                     /label="FS3_CYCtt_FS4"
     primer          <7219..>7222
                     /label="5_BsaI_FsC_Xho"
     FS              7219..7222
                     /label="FS 4"
     primer          complement(7223..>7226)
                     /label="6_BsaI_FsC"
     promoter        7231..7574
                     /label="TEFpromoter"
     CDS             7575..8603
                     /label="hphR (Hygromycin)"
     terminator      8612..8806
                     /label="TEFterminator"
     misc_feature    8811..9322
                     /label="CEN6_ARS4"
     mutation        9032
                     /label="G/A"
ORIGIN
        1 CGCTGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT
       61 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT
      121 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC
      181 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA
      241 GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT
      301 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA
      361 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG
      421 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC
      481 CTAACTACGG CTACACTAGA AGAACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA
      541 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG
      601 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT
      661 TGATCTTTTC TACGGGGTGG AGcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcGATCGGAG GATCCTTTTT TGTAGAAATG TCTTGGTGTC CTCGTCCAAT CAGGTAGCCA
      781 TCTCTGAAAT ATCTGGCTCC GTTGCAACTC CGAACGACCT GCTGGCAACG TAAAATTCTC
      841 CGGGGTAAAA CTTAAATGTG GAGTAATGGA ACCAGAAACG TCTCTTCCCT TCTCTCTCCT
      901 TCCACCGCCC GTTACCGTCC CTAGGAAATT TTACTCTGCT GGAGAGCTTC TTCTACGGCC
      961 CCCTTGCAGC AATGCTCTTC CCAGCATTAC GTTGCGGGTA AAACGGAGGT CGTGTACCCG
     1021 ACCTAGCAGC CCAGGGATGG AAAAGTCCCG GCCGTCGCTG GCAATAATAG CGGGCGGACG
     1081 CATGTCATGA GATTATTGGA AACCACCAGA ATCGAATATA AAAGGCGAAC ACCTTTCCCA
     1141 ATTTTGGTTT CTCCTGACCC AAAGACTTTA AATTTAATTT ATTTGTCCCT ATTTCAATCA
     1201 ATTGAACAAC TATCAAAACA CCATGGCGTC TTCGGATCCC TCGAGGAAGA CGCGCTTTTA
     1261 GTGTACATCT GATAATATAG TACTACCACG TATGATAATG TAGAGAATAG GCTTCCTTGT
     1321 CGAGTGTGTT TGCAGTTTTC TTGAGTTTCA AGGTTTAAAT GCTGGTATAT TAGTTCATCG
     1381 AAGGTTTCAG CCAATAGCAC CTTAAATCAA TCAAACTAAT TCGACTCTTA CGAAAGAGCC
     1441 TACTGTGTTT AGTATCGAAG TCGTTTACCT TTCATGTTGA ATAGCTTCCT CTCTGACCCT
     1501 AACATTTCAA GATCCTCCTA AAGTTACCCG GATTGTGAAA TTCTAATGAT CCACCTGCCC
     1561 AATGCATTTT TTCTTTATTC AGTTTACCTT TTTTACCTAA TATACGAGCT TGTTAAAGTA
     1621 AGTGGCACTG CAATACTAGG CTTATTGTTG ATATTATGAT GAATCGTTTT CACAAACTTG
     1681 ATTTCCTGTG AACTCACCAT GTACTAAGGA AAAAAACATG CATCACCATC TGAATATTTG
     1741 ACCGCTCCGG GGAGGGAAGA ATATGAACCA CGATTGAAAG ACTTTGGGTT TGCCGATTCT
     1801 AAAACACTTA CTGAACAGAA AAGAAGTGAA TTATTAGAGA GTTTATGCAA CGGAGATCAT
     1861 GAGCTTAGAA AGAACGTAGG CTGGGCAACA ACATCCATTT CTGCCTGCGA TATTTCCAGT
     1921 GCAATGCTAA AGCCACATAA TATAGGCAAT TACAACTTGA ATGAGCAGGC TCATGATGTC
     1981 ACCATGGAAC TGATAGATAA AGTCATCCAA AAAGGTGTCA ATTTACGGCA CGTCTACGTA
     2041 GATACTGTCG GACCTCCAGC ATCATACCAA AAGAAACTGC AACAAAGGTT TCCCGCACTG
     2101 CTGATCACCG TATCTAAAAA AGCAGATGCC ATCTATCCAT GTGTTTCAAG TGCTTCAGTT
     2161 TGCGCCAAGG TCACTAGAGA CGTCTGTTTG AGAACCTCAA ATGGGGAGGA GAACCTATTA
     2221 GATTGTGGGT CTGGATATCC CTCGGACCCA AATACTAAGA AATGGCTAGC CCAGTCTATA
     2281 CATCCATTGT TTGGCTGGAG TCGTGATGTC AGGTTTTCGT GGCAGACGGC CAAAGATTGT
     2341 CTTGAGAGAC ACAACTCAAT CAAAGTTGTG TTTGGCGAAG AGGAGCCTAC CCCGGTTAAT
     2401 TTCGGTGATC TGGGGTTGAG CAAACAGAAG CCCGACTTTA AGGATTTAGA AATCAGTACC
     2461 AGTTGGTGGG GACATTAATT GGTGGAGGAG AGCCAATATT CTCAACTGTT CGTAGGACCA
     2521 TAACCCCCCA CTCGCACCAG AAATACTCAT TGGCGCTGAC AACTAGTTTG AGCCGCTTCA
     2581 ATTGCGGCAA GTCCCCAGTT TTGTTTGGTA CTCCCCTCTC GCCTCGCGTG TTTTTGTTCC
     2641 TCCGAATTGT GCATCACCCT GTCACATCCG TACACCATCG GAACACTCTG CTCCTCCTCC
     2701 TTTCTCCTCT TAATTGAACG TATATCTAAT TACCCTAATA ACAAGTATCT TCAACCATGG
     2761 ACAAGAAGTA CTCCATTGGG CTCGATATCG GCACAAACAG CGTCGGTTGG GCCGTCATTA
     2821 CGGACGAGTA CAAGGTGCCG AGCAAAAAAT TCAAAGTTCT GGGCAATACC GATCGCCACA
     2881 GCATAAAGAA GAACCTCATT GGCGCCCTCC TGTTCGACTC CGGGGAGACG GCCGAAGCCA
     2941 CGCGGCTCAA AAGAACAGCA CGGCGCAGAT ATACCCGCAG AAAGAATCGG ATCTGCTACC
     3001 TGCAGGAGAT CTTTAGTAAT GAGATGGCTA AGGTGGATGA CTCTTTCTTC CATAGGCTGG
     3061 AGGAGTCCTT TTTGGTGGAG GAGGATAAAA AGCACGAGCG CCACCCAATC TTTGGCAATA
     3121 TCGTGGACGA GGTGGCGTAC CATGAAAAGT ACCCAACCAT ATATCATCTG AGGAAGAAGC
     3181 TTGTAGACAG TACTGATAAG GCTGACTTGC GGTTGATCTA TCTCGCGCTG GCGCATATGA
     3241 TCAAATTTCG GGGACACTTC CTCATCGAGG GGGACCTGAA CCCAGACAAC AGCGATGTCG
     3301 ACAAACTCTT TATCCAACTG GTTCAGACTT ACAATCAGCT TTTCGAAGAG AACCCGATCA
     3361 ACGCATCCGG AGTTGACGCC AAAGCAATCC TGAGCGCTAG GCTGTCCAAA TCCCGGCGGC
     3421 TCGAAAACCT CATCGCACAG CTCCCTGGGG AGAAGAAGAA CGGCCTGTTT GGTAATCTTA
     3481 TCGCCCTGTC ACTCGGGCTG ACCCCCAACT TTAAATCTAA CTTCGACCTG GCCGAAGATG
     3541 CCAAGCTTCA ACTGAGCAAA GACACCTACG ATGATGATCT CGACAATCTG CTGGCCCAGA
     3601 TCGGCGACCA GTACGCAGAC CTTTTTTTGG CGGCAAAGAA CCTGTCAGAC GCCATTCTGC
     3661 TGAGTGATAT TCTGCGAGTG AACACGGAGA TCACCAAAGC TCCGCTGAGC GCTAGTATGA
     3721 TCAAGCGCTA TGATGAGCAC CACCAAGACT TGACTTTGCT GAAGGCCCTT GTCAGACAGC
     3781 AACTGCCTGA GAAGTACAAG GAAATTTTCT TCGATCAGTC TAAAAATGGC TACGCCGGAT
     3841 ACATTGACGG CGGAGCAAGC CAGGAGGAAT TTTACAAATT TATTAAGCCC ATCTTGGAAA
     3901 AAATGGACGG CACCGAGGAG CTGCTGGTAA AGCTTAACAG AGAAGATCTG TTGCGCAAAC
     3961 AGCGCACTTT CGACAATGGA AGCATCCCCC ACCAGATTCA CCTGGGCGAA CTGCACGCTA
     4021 TCCTCAGGCG GCAAGAGGAT TTCTACCCCT TTTTGAAAGA TAACAGGGAA AAGATTGAGA
     4081 AAATCCTCAC ATTTCGGATA CCCTACTATG TAGGCCCCCT CGCCCGGGGA AATTCCAGAT
     4141 TCGCGTGGAT GACTCGCAAA TCAGAAGAGA CtATCACTCC CTGGAACTTC GAGGAAGTCG
     4201 TGGATAAGGG GGCCTCTGCC CAGTCCTTCA TCGAAAGGAT GACTAACTTT GATAAAAATC
     4261 TGCCTAACGA AAAGGTGCTT CCTAAACACT CTCTGCTGTA CGAGTACTTC ACAGTTTATA
     4321 ACGAGCTCAC CAAGGTCAAA TACGTCACAG AAGGGATGAG AAAGCCAGCA TTCCTGTCTG
     4381 GAGAGCAGAA GAAAGCTATC GTGGACCTCC TCTTCAAGAC GAACCGGAAA GTTACCGTGA
     4441 AACAGCTCAA AGAgGACTAT TTCAAAAAGA TTGAATGTTT CGACTCTGTT GAAATCAGCG
     4501 GAGTGGAGGA TCGCTTCAAC GCATCCCTGG GAACGTATCA CGATCTCCTG AAAATCATTA
     4561 AAGACAAGGA CTTCCTGGAC AATGAGGAGA ACGAGGACAT TCTTGAGGAC ATTGTCCTCA
     4621 CCCTTACGTT GTTTGAAGAT AGGGAGATGA TTGAAGAACG CTTGAAAACT TACGCTCATC
     4681 TCTTCGACGA CAAAGTCATG AAACAGCTCA AGAGGCGCCG ATATACAGGA TGGGGGCGGC
     4741 TGTCAAGAAA ACTGATCAAT GGGATCCGAG ACAAGCAGAG TGGAAAGACA ATCCTGGATT
     4801 TTCTTAAGTC CGATGGATTT GCCAACCGGA ACTTCATGCA GTTGATCCAT GATGACTCTC
     4861 TCACCTTTAA GGAGGACATC CAGAAAGCAC AAGTTTCTGG CCAGGGGGAC AGcCTTCACG
     4921 AGCACATCGC TAATCTTGCA GGTAGCCCAG CTATCAAAAA GGGAATACTG CAGACCGTTA
     4981 AGGTCGTGGA TGAACTCGTC AAAGTAATGG GAAGGCATAA GCCCGAGAAT ATCGTTATCG
     5041 AGATGGCCCG AGAGAACCAA ACTACCCAGA AGGGACAGAA GAACAGTAGG GAAAGGATGA
     5101 AGAGGATTGA AGAGGGTATA AAAGAACTGG GGTCCCAAAT CCTTAAGGAA CACCCAGTTG
     5161 AAAACACCCA GCTTCAGAAT GAGAAGCTCT ACCTGTACTA CCTGCAGAAC GGCAGGGACA
     5221 TGTACGTGGA TCAGGAACTG GACATCAATC GGCTCTCCGA CTACGACGTG GATCATATCG
     5281 TGCCCCAGTC TTTTCTCAAA GATGATTCTA TTGATAATAA AGTGTTGACA AGATCCGATA
     5341 AAAATAGAGG GAAGAGTGAT AACGTCCCCT CAGAAGAAGT TGTCAAGAAA ATGAAAAATT
     5401 ATTGGCGGCA GCTGCTGAAC GCCAAACTGA TCACACAACG GAAGTTCGAT AATCTGACTA
     5461 AGGCTGAACG AGGTGGCCTG TCTGAGTTGG ATAAAGCCGG CTTCATCAAA AGGCAGCTTG
     5521 TTGAGACACG CCAGATCACC AAGCACGTGG CCCAAATTCT CGATTCACGC ATGAACACCA
     5581 AGTACGATGA AAATGACAAA CTGATTCGAG AGGTGAAAGT TATTACTCTG AAGTCTAAGC
     5641 TcGTCTCAGA TTTCAGAAAG GACTTTCAGT TTTATAAGGT GAGAGAGATC AACAATTACC
     5701 ACCATGCGCA TGATGCCTAC CTGAATGCAG TGGTAGGCAC TGCACTTATC AAAAAATATC
     5761 CCAAGCTTGA ATCTGAATTT GTTTACGGAG ACTATAAAGT GTACGATGTT AGGAAAATGA
     5821 TCGCAAAGTC TGAGCAGGAA ATAGGCAAGG CCACCGCTAA GTACTTCTTT TACAGCAATA
     5881 TTATGAATTT TTTCAAGACC GAGATTACAC TGGCCAATGG AGAGATTCGG AAGCGACCAC
     5941 TTATCGAAAC AAACGGAGAA ACAGGAGAAA TCGTGTGGGA CAAGGGTAGG GATTTCGCGA
     6001 CAGTCCGGAA GGTCCTGTCC ATGCCGCAGG TGAACATCGT TAAAAAGACC GAAGTACAGA
     6061 CCGGAGGCTT CTCCAAGGAA AGTATCCTCC CGAAAAGGAA CAGCGACAAG CTGATCGCAC
     6121 GCAAAAAAGA TTGGGACCCC AAGAAATACG GCGGATTCGA TTCTCCTACA GTCGCTTACA
     6181 GTGTACTGGT TGTGGCCAAA GTGGAGAAAG GGAAGTCTAA AAAACTCAAA AGCGTCAAGG
     6241 AACTGCTGGG CATCACAATC ATGGAGCGAT CAAGCTTCGA AAAAAACCCC ATCGACTTTC
     6301 TCGAGGCGAA AGGATATAAA GAGGTCAAAA AAGACCTCAT CATTAAGCTT CCCAAGTACT
     6361 CTCTCTTTGA GCTTGAAAAC GGCCGGAAAC GAATGCTCGC TAGTGCGGGC GAGCTGCAGA
     6421 AAGGTAACGA GCTGGCACTG CCCTCTAAAT ACGTTAATTT CTTGTATCTG GCCAGCCACT
     6481 ATGAAAAGCT CAAAGGGTCc CCCGAAGATA ATGAGCAGAA GCAGCTGTTC GTGGAACAAC
     6541 ACAAACACTA CCTTGATGAG ATCATCGAGC AAATAAGCGA ATTCTCCAAA AGAGTGATCC
     6601 TCGCCGACGC TAACCTCGAT AAGGTGCTTT CTGCTTACAA TAAGCACAGG GATAAGCCCA
     6661 TCAGGGAGCA GGCAGAAAAC ATTATCCACT TGTTTACTCT GACCAACTTG GGCGCGCCTG
     6721 CAGCCTTCAA GTACTTCGAC ACCACCATAG ACAGAAAGCG GTACACCTCT ACAAAGGAGG
     6781 TCCTGGACGC CACACTGATT CATCAGTCAA TTACGGGGCT CTATGAAACA AGAATCGACC
     6841 TCTCTCAGCT CGGTGGAGAC AGCAGGGCTG ACCCCAAGAA GAAGAGGAAG GTGTGAGCTT
     6901 CGTCCGACGG CGGCCCACGG GTCCCAGGCC TCGGAGATCC GTCCCCCTTT TCCTTTGTCG
     6961 ATATCATGTA ATTAGTTATG TCACGCTTAC ATTCACGCCC TCCCCCCACA TCCGCTCTAA
     7021 CCGAAAAGGA AGGAGTTAGA CAACCTGAAG TCTAGGTCCC TATTTATTTT TTTATAGTTA
     7081 TGTTAGTATT AAGAACGTTA TTTATATTTC AAATTTTTCT TTTTTTTCTG TACAGACGCG
     7141 TGTACGCATG TAACATTATA CTGAAAACCT TGCTTGAGAA GGTTTTGGGA CGCTCGAAGG
     7201 CTTTAATTTG CAAGCTGGCG CTAATTCATG GACATGGAGG CCCAGAATAC CCTCCTTGAC
     7261 AGTCTTGACG TGCGCAGCTC AGGGGCATGA TGTGACTGTC GCCCGTACAT TTAGCCCATA
     7321 CATCCCCATG TATAATCATT TGCATCCATA CATTTTGATG GCCGCACGGC GCGAAGCAAA
     7381 AATTACGGCT CCTCGCTGCA GACGTGCGAG CAGGGAAACG CTCCCCTCAC AGACGCGTTG
     7441 AATTGTCCCC ACGCCGCGCC CCTGTAGAGA AATATAAAAG GTTAGGATTT GCCACTGAGG
     7501 TTCTTCTTTC ATATACTTCC TTTTAAAATC TTGCTAGGAT ACAGTTCTCA CATCACATCC
     7561 GAACATAAAC AACCATGGGT AAAAAGCCTG AACTCACCGC GACGTCTGTC GAGAAGTTTC
     7621 TGATCGAAAA GTTCGACAGC GTCTCCGACC TGATGCAGCT CTCGGAGGGC GAAGAATCTC
     7681 GTGCTTTCAG CTTCGATGTA GGAGGGCGTG GATATGTCCT GCGGGTAAAT AGCTGCGCCG
     7741 ATGGTTTCTA CAAAGATCGT TATGTTTATC GGCACTTTGC ATCGGCCGCG CTCCCGATTC
     7801 CGGAAGTGCT TGACATTGGG GAATTCAGCG AGAGCCTGAC CTATTGCATC TCCCGCCGTG
     7861 CACAGGGTGT CACGTTGCAA GACCTCCCTG AAACCGAACT GCCCGCTGTT CTGCAGCCGG
     7921 TCGCGGAGGC CATGGATGCG ATCGCTGCGG CCGATCTTAG CCAGACGAGC GGGTTCGGCC
     7981 CATTCGGACC GCAAGGAATC GGTCAATACA CTACATGGCG TGATTTCATA TGCGCGATTG
     8041 CTGATCCCCA TGTGTATCAC TGGCAAACTG TGATGGACGA CACCGTCAGT GCGTCCGTCG
     8101 CGCAGGCTCT CGATGAGCTG ATGCTTTGGG CCGAGGACTG CCCCGAAGTC CGGCACCTCG
     8161 TGCACGCGGA TTTCGGCTCC AACAATGTCC TGACGGACAA TGGCCGCATA ACAGCGGTCA
     8221 TTGACTGGAG CGAGGCGATG TTCGGGGATT CCCAATACGA GGTCGCCAAC ATCTTCTTCT
     8281 GGAGGCCGTG GTTGGCTTGT ATGGAGCAGC AGACGCGCTA CTTCGAGCGG AGGCATCCGG
     8341 AGCTTGCAGG ATCGCCGCGG CTCCGGGCGT ATATGCTCCG CATTGGTCTT GACCAACTCT
     8401 ATCAGAGCTT GGTTGACGGC AATTTCGATG ATGCAGCTTG GGCGCAGGGT CGATGCGACG
     8461 CAATCGTCCG ATCCGGAGCC GGGACTGTCG GGCGTACACA AATCGCCCGC AGAAGCGCGG
     8521 CCGTCTGGAC CGATGGCTGT GTAGAAGTAC TCGCCGATAG TGGAAACCGA CGCCCCAGCA
     8581 CTCGTCCGAG GGCAAAGGAA TAATCAGTAC TGACAATAAA AAGATTCTTG TTTTCAAGAA
     8641 CTTGTCATTT GTATAGTTTT TTTATATTGT AGTTGTTCTA TTTTAATCAA ATGTTAGCGT
     8701 GATTTATATT TTTTTTCGCC TCGACATCAT CTGCCCAGAT GCGAAGTTAA GTGCGCAGAA
     8761 AGTAATATCA TGCGTCAATC GTATGTGAAT GCTGGTCGCT ATACTGGCTT CCTTTTCATC
     8821 ACGTGCTATA AAAATAATTA TAATTTAAAT TTTTTAATAT AAATATATAA ATTAAAAATA
     8881 GAAAGTAAAA AAAGAAATTA AAGAAAAAAT AGTTTTTGTT TTCCGAAGAT GTAAAAGACT
     8941 CTAGGGGGAT CGCCAACAAA TACTACCTTT TATCTTGCTC TTCCTGCTCT CAGGTATTAA
     9001 TGCCGAATTG TTTCATCTTG TCTGTGTAGA ACACCACACA CGAAAATCCT GTGATTTTAC
     9061 ATTTTACTTA TCGTTAATCG AATGTATATC TATTTAATCT GCTTTTCTTG TCTAATAAAT
     9121 ATATATGTAA AGTACGCTTT TTGTTGAAAT TTTTTAAACC TTTGTTTATT TTTTTTTCTT
     9181 CATTCCGTAA CTCTTCTACC TTCTTTATTT ACTTTCTAAA ATCCAAATAC AAAACATAAA
     9241 AATAAATAAA CACAGAGTAA ATTCCCAAAT TATTCCATCA TTAAAAGATA CGAGGCGCGT
     9301 GTAAGTTACA GGCAAGCGAT CG
//
LOCUS       BB3_pGAP_Linker_BbsI(intern)_RPS25Att_pPFK300_Cas9_cycTT_G418      8326 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Vector          join(6454..8326,1..730)
                     /origin="BB3_pTEF_KanR_TEFt_AC (2,633bp circular)Target
                     vector: from BsaI cut at 760^761 to BsaI cut at 730^731,
                     2,603bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     rep_origin      complement(13..686)
                     /label="pUC_ORI_aus_pGAPZB"
     primer          448..467
                     /label="J 77"
     primer          524..541
                     /label="B1_13"
     terminator      691..730
                     /label="BBa_B1007"
     FS              731..734
                     /label="FS A"
     Vector          join(1751..>1754,<731..734)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 734^735
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        737..>1229
                     /label="pGAP"
     Restriction_sit join(1229^1230,complement(1229^1230))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1230..>1261
                     /label="L_FS23_BbsI_fw"
     misc_feature    1230..1261
                     /feature_type="Insertion"
                     /origin="PCR_Linker_FS23_BbsI(intern) (54bp
                     linear)Fragment: from BpiI cut at 9^10 to BpiI cut at
                     41^42, 32bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1230..1233
                     /label="FS2"
     primer          complement(<1234..>1265)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1262..1294,complement(1295..1727),complement(1728..>1754))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1266..>1750
                     /note="Forward primer name: RPS25ATT_M1_FW"
                     /note="Forward primer: GacagGtcTccgCTTCCTTGTCGAGTGTGTTT"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_RV"
                     /note="Reverse primer:
                     gatcggtctCcAgctGTCAAATATTCAGATGGTGA"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 458"
                     /label="rps25aTT"
     primer          1319..1339
                     /label="J_89"
     FS              1751..1754
                     /label="FS 4"
     primer          <1755..>1758
                     /label="3_BsaI_FsB_Bam"
     misc_feature    join(<6450..>6453,<1755..>1758)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (108bp linear)Fragment: full
                     sequenceOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1755..1758
                     /label="FS B"
     Vector          join(6450..>6453,<1755..1758)
                     /origin="BB2_L_BC_syn_BbsI_7 (2,215bp circular)Target
                     vector: from BpiI cut at 639^640 to BpiI cut at 607^608,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          complement(<1759..>1762)
                     /label="4_BsaI_FsB_BamXho"
     primer          1766..1790
                     /label="J_103"
     promoter        1771..1986
                     /label="pPpPfk"
     primer          <1987..2011
                     /label="FS2_Cas9_BsaI_fw"
     CDS             1988..6127
                     /label="Cas9"
     Taming          2038
                     /label="C>T"
     primer          complement(3399..3418)
                     /label="1_cas9_rev"
     primer          3399..3418
                     /label="1_cas9_fw"
     Taming          3403
                     /label="C>t"
     primer          complement(3673..3694)
                     /label="2_cas9_rev"
     primer          3673..3694
                     /label="2_cas9_fw"
     Taming          3685
                     /label="A>g"
     primer          complement(4142..4159)
                     /label="3_cas9_rev"
     primer          4142..4159
                     /label="3_cas9_fw"
     Taming          4144
                     /label="T>c"
     primer          complement(4857..4878)
                     /label="4_cas9_rev"
     primer          4857..4878
                     /label="4_cas9_fw"
     Taming          4873
                     /label="G>c"
     primer          complement(5724..5741)
                     /label="5_cas9_rev"
     primer          5724..5742
                     /label="5_cas9_fw"
     Taming          5731
                     /label="T>c"
     NLS             6092..6127
                     /label="NLS SV40"
     primer          complement(6105..>6131)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6128..>6449
                     /label="FS3_CYCtt_FS4"
     primer          <6450..>6453
                     /label="5_BsaI_FsC_Xho"
     FS              6450..6453
                     /label="FS 4"
     primer          complement(6454..>6457)
                     /label="6_BsaI_FsC"
     promoter        6462..6805
                     /label="pTEF (Ashbya gossypii)"
     mutation        6635
                     /label="C to G"
     primer          complement(6689..6708)
                     /label="J_78"
     CDS             6806..7615
                     /label="KanR / G418"
     terminator      7624..7822
                     /label="TEFtt (Ashbya gossypii)"
     misc_feature    7823..8326
                     /label="CEN6_ARS4"
     mutation        8044
                     /label="G/A, actually G/C"
ORIGIN
        1 AGCGATCGCG CTGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT
       61 GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG
      121 TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC
      181 CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC
      241 TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT
      301 CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT
      361 ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC
      421 AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA
      481 GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG CTCTGCTGAA
      541 GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG
      601 TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA
      661 AGATCCTTTG ATCTTTTCTA CGGGGTGGAG cgcaaaaaac cccgcccctg acagggcggg
      721 gttttttcgc GATCGGAGGA TCCTTTTTTG TAGAAATGTC TTGGTGTCCT CGTCCAATCA
      781 GGTAGCCATC TCTGAAATAT CTGGCTCCGT TGCAACTCCG AACGACCTGC TGGCAACGTA
      841 AAATTCTCCG GGGTAAAACT TAAATGTGGA GTAATGGAAC CAGAAACGTC TCTTCCCTTC
      901 TCTCTCCTTC CACCGCCCGT TACCGTCCCT AGGAAATTTT ACTCTGCTGG AGAGCTTCTT
      961 CTACGGCCCC CTTGCAGCAA TGCTCTTCCC AGCATTACGT TGCGGGTAAA ACGGAGGTCG
     1021 TGTACCCGAC CTAGCAGCCC AGGGATGGAA AAGTCCCGGC CGTCGCTGGC AATAATAGCG
     1081 GGCGGACGCA TGTCATGAGA TTATTGGAAA CCACCAGAAT CGAATATAAA AGGCGAACAC
     1141 CTTTCCCAAT TTTGGTTTCT CCTGACCCAA AGACTTTAAA TTTAATTTAT TTGTCCCTAT
     1201 TTCAATCAAT TGAACAACTA TCAAAACACC ATGGCGTCTT CGGATCCCTC GAGGAAGACG
     1261 CGCTTTTAGT GTACATCTGA TAATATAGTA CTACCACGTA TGATAATGTA GAGAATAGGC
     1321 TTCCTTGTCG AGTGTGTTTG CAGTTTTCTT GAGTTTCAAG GTTTAAATGC TGGTATATTA
     1381 GTTCATCGAA GGTTTCAGCC AATAGCACCT TAAATCAATC AAACTAATTC GACTCTTACG
     1441 AAAGAGCCTA CTGTGTTTAG TATCGAAGTC GTTTACCTTT CATGTTGAAT AGCTTCCTCT
     1501 CTGACCCTAA CATTTCAAGA TCCTCCTAAA GTTACCCGGA TTGTGAAATT CTAATGATCC
     1561 ACCTGCCCAA TGCATTTTTT CTTTATTCAG TTTACCTTTT TTACCTAATA TACGAGCTTG
     1621 TTAAAGTAAG TGGCACTGCA ATACTAGGCT TATTGTTGAT ATTATGATGA ATCGTTTTCA
     1681 CAAACTTGAT TTCCTGTGAA CTCACCATGT ACTAAGGAAA AAAACATGCA TCACCATCTG
     1741 AATATTTGAC CGCTCCGGGG AGCCTGCCAT TAGGCAAACA CCAACTAGGA ACCTACACCA
     1801 AGTTATGAAG GAGCTTGGAA AGGATTATAA AAAATGCGAT TACCTAATAC ACACACGATG
     1861 CCTGAGCGGT TAATGTGAAA TTTCCCGGCC ATCGATCGCC CCACAATCAG ACTATTTTCG
     1921 CTCATACAAT AAAAAGTCTC CTGTTTCCAC CCGAGTATCA CCAGAACAAA GTAACAAATA
     1981 CAAAACCATG GACAAGAAGT ACTCCATTGG GCTCGATATC GGCACAAACA GCGTCGGTTG
     2041 GGCCGTCATT ACGGACGAGT ACAAGGTGCC GAGCAAAAAA TTCAAAGTTC TGGGCAATAC
     2101 CGATCGCCAC AGCATAAAGA AGAACCTCAT TGGCGCCCTC CTGTTCGACT CCGGGGAGAC
     2161 GGCCGAAGCC ACGCGGCTCA AAAGAACAGC ACGGCGCAGA TATACCCGCA GAAAGAATCG
     2221 GATCTGCTAC CTGCAGGAGA TCTTTAGTAA TGAGATGGCT AAGGTGGATG ACTCTTTCTT
     2281 CCATAGGCTG GAGGAGTCCT TTTTGGTGGA GGAGGATAAA AAGCACGAGC GCCACCCAAT
     2341 CTTTGGCAAT ATCGTGGACG AGGTGGCGTA CCATGAAAAG TACCCAACCA TATATCATCT
     2401 GAGGAAGAAG CTTGTAGACA GTACTGATAA GGCTGACTTG CGGTTGATCT ATCTCGCGCT
     2461 GGCGCATATG ATCAAATTTC GGGGACACTT CCTCATCGAG GGGGACCTGA ACCCAGACAA
     2521 CAGCGATGTC GACAAACTCT TTATCCAACT GGTTCAGACT TACAATCAGC TTTTCGAAGA
     2581 GAACCCGATC AACGCATCCG GAGTTGACGC CAAAGCAATC CTGAGCGCTA GGCTGTCCAA
     2641 ATCCCGGCGG CTCGAAAACC TCATCGCACA GCTCCCTGGG GAGAAGAAGA ACGGCCTGTT
     2701 TGGTAATCTT ATCGCCCTGT CACTCGGGCT GACCCCCAAC TTTAAATCTA ACTTCGACCT
     2761 GGCCGAAGAT GCCAAGCTTC AACTGAGCAA AGACACCTAC GATGATGATC TCGACAATCT
     2821 GCTGGCCCAG ATCGGCGACC AGTACGCAGA CCTTTTTTTG GCGGCAAAGA ACCTGTCAGA
     2881 CGCCATTCTG CTGAGTGATA TTCTGCGAGT GAACACGGAG ATCACCAAAG CTCCGCTGAG
     2941 CGCTAGTATG ATCAAGCGCT ATGATGAGCA CCACCAAGAC TTGACTTTGC TGAAGGCCCT
     3001 TGTCAGACAG CAACTGCCTG AGAAGTACAA GGAAATTTTC TTCGATCAGT CTAAAAATGG
     3061 CTACGCCGGA TACATTGACG GCGGAGCAAG CCAGGAGGAA TTTTACAAAT TTATTAAGCC
     3121 CATCTTGGAA AAAATGGACG GCACCGAGGA GCTGCTGGTA AAGCTTAACA GAGAAGATCT
     3181 GTTGCGCAAA CAGCGCACTT TCGACAATGG AAGCATCCCC CACCAGATTC ACCTGGGCGA
     3241 ACTGCACGCT ATCCTCAGGC GGCAAGAGGA TTTCTACCCC TTTTTGAAAG ATAACAGGGA
     3301 AAAGATTGAG AAAATCCTCA CATTTCGGAT ACCCTACTAT GTAGGCCCCC TCGCCCGGGG
     3361 AAATTCCAGA TTCGCGTGGA TGACTCGCAA ATCAGAAGAG ACtATCACTC CCTGGAACTT
     3421 CGAGGAAGTC GTGGATAAGG GGGCCTCTGC CCAGTCCTTC ATCGAAAGGA TGACTAACTT
     3481 TGATAAAAAT CTGCCTAACG AAAAGGTGCT TCCTAAACAC TCTCTGCTGT ACGAGTACTT
     3541 CACAGTTTAT AACGAGCTCA CCAAGGTCAA ATACGTCACA GAAGGGATGA GAAAGCCAGC
     3601 ATTCCTGTCT GGAGAGCAGA AGAAAGCTAT CGTGGACCTC CTCTTCAAGA CGAACCGGAA
     3661 AGTTACCGTG AAACAGCTCA AAGAgGACTA TTTCAAAAAG ATTGAATGTT TCGACTCTGT
     3721 TGAAATCAGC GGAGTGGAGG ATCGCTTCAA CGCATCCCTG GGAACGTATC ACGATCTCCT
     3781 GAAAATCATT AAAGACAAGG ACTTCCTGGA CAATGAGGAG AACGAGGACA TTCTTGAGGA
     3841 CATTGTCCTC ACCCTTACGT TGTTTGAAGA TAGGGAGATG ATTGAAGAAC GCTTGAAAAC
     3901 TTACGCTCAT CTCTTCGACG ACAAAGTCAT GAAACAGCTC AAGAGGCGCC GATATACAGG
     3961 ATGGGGGCGG CTGTCAAGAA AACTGATCAA TGGGATCCGA GACAAGCAGA GTGGAAAGAC
     4021 AATCCTGGAT TTTCTTAAGT CCGATGGATT TGCCAACCGG AACTTCATGC AGTTGATCCA
     4081 TGATGACTCT CTCACCTTTA AGGAGGACAT CCAGAAAGCA CAAGTTTCTG GCCAGGGGGA
     4141 CAGcCTTCAC GAGCACATCG CTAATCTTGC AGGTAGCCCA GCTATCAAAA AGGGAATACT
     4201 GCAGACCGTT AAGGTCGTGG ATGAACTCGT CAAAGTAATG GGAAGGCATA AGCCCGAGAA
     4261 TATCGTTATC GAGATGGCCC GAGAGAACCA AACTACCCAG AAGGGACAGA AGAACAGTAG
     4321 GGAAAGGATG AAGAGGATTG AAGAGGGTAT AAAAGAACTG GGGTCCCAAA TCCTTAAGGA
     4381 ACACCCAGTT GAAAACACCC AGCTTCAGAA TGAGAAGCTC TACCTGTACT ACCTGCAGAA
     4441 CGGCAGGGAC ATGTACGTGG ATCAGGAACT GGACATCAAT CGGCTCTCCG ACTACGACGT
     4501 GGATCATATC GTGCCCCAGT CTTTTCTCAA AGATGATTCT ATTGATAATA AAGTGTTGAC
     4561 AAGATCCGAT AAAAATAGAG GGAAGAGTGA TAACGTCCCC TCAGAAGAAG TTGTCAAGAA
     4621 AATGAAAAAT TATTGGCGGC AGCTGCTGAA CGCCAAACTG ATCACACAAC GGAAGTTCGA
     4681 TAATCTGACT AAGGCTGAAC GAGGTGGCCT GTCTGAGTTG GATAAAGCCG GCTTCATCAA
     4741 AAGGCAGCTT GTTGAGACAC GCCAGATCAC CAAGCACGTG GCCCAAATTC TCGATTCACG
     4801 CATGAACACC AAGTACGATG AAAATGACAA ACTGATTCGA GAGGTGAAAG TTATTACTCT
     4861 GAAGTCTAAG CTcGTCTCAG ATTTCAGAAA GGACTTTCAG TTTTATAAGG TGAGAGAGAT
     4921 CAACAATTAC CACCATGCGC ATGATGCCTA CCTGAATGCA GTGGTAGGCA CTGCACTTAT
     4981 CAAAAAATAT CCCAAGCTTG AATCTGAATT TGTTTACGGA GACTATAAAG TGTACGATGT
     5041 TAGGAAAATG ATCGCAAAGT CTGAGCAGGA AATAGGCAAG GCCACCGCTA AGTACTTCTT
     5101 TTACAGCAAT ATTATGAATT TTTTCAAGAC CGAGATTACA CTGGCCAATG GAGAGATTCG
     5161 GAAGCGACCA CTTATCGAAA CAAACGGAGA AACAGGAGAA ATCGTGTGGG ACAAGGGTAG
     5221 GGATTTCGCG ACAGTCCGGA AGGTCCTGTC CATGCCGCAG GTGAACATCG TTAAAAAGAC
     5281 CGAAGTACAG ACCGGAGGCT TCTCCAAGGA AAGTATCCTC CCGAAAAGGA ACAGCGACAA
     5341 GCTGATCGCA CGCAAAAAAG ATTGGGACCC CAAGAAATAC GGCGGATTCG ATTCTCCTAC
     5401 AGTCGCTTAC AGTGTACTGG TTGTGGCCAA AGTGGAGAAA GGGAAGTCTA AAAAACTCAA
     5461 AAGCGTCAAG GAACTGCTGG GCATCACAAT CATGGAGCGA TCAAGCTTCG AAAAAAACCC
     5521 CATCGACTTT CTCGAGGCGA AAGGATATAA AGAGGTCAAA AAAGACCTCA TCATTAAGCT
     5581 TCCCAAGTAC TCTCTCTTTG AGCTTGAAAA CGGCCGGAAA CGAATGCTCG CTAGTGCGGG
     5641 CGAGCTGCAG AAAGGTAACG AGCTGGCACT GCCCTCTAAA TACGTTAATT TCTTGTATCT
     5701 GGCCAGCCAC TATGAAAAGC TCAAAGGGTC cCCCGAAGAT AATGAGCAGA AGCAGCTGTT
     5761 CGTGGAACAA CACAAACACT ACCTTGATGA GATCATCGAG CAAATAAGCG AATTCTCCAA
     5821 AAGAGTGATC CTCGCCGACG CTAACCTCGA TAAGGTGCTT TCTGCTTACA ATAAGCACAG
     5881 GGATAAGCCC ATCAGGGAGC AGGCAGAAAA CATTATCCAC TTGTTTACTC TGACCAACTT
     5941 GGGCGCGCCT GCAGCCTTCA AGTACTTCGA CACCACCATA GACAGAAAGC GGTACACCTC
     6001 TACAAAGGAG GTCCTGGACG CCACACTGAT TCATCAGTCA ATTACGGGGC TCTATGAAAC
     6061 AAGAATCGAC CTCTCTCAGC TCGGTGGAGA CAGCAGGGCT GACCCCAAGA AGAAGAGGAA
     6121 GGTGTGAGCT TCGTCCGACG GCGGCCCACG GGTCCCAGGC CTCGGAGATC CGTCCCCCTT
     6181 TTCCTTTGTC GATATCATGT AATTAGTTAT GTCACGCTTA CATTCACGCC CTCCCCCCAC
     6241 ATCCGCTCTA ACCGAAAAGG AAGGAGTTAG ACAACCTGAA GTCTAGGTCC CTATTTATTT
     6301 TTTTATAGTT ATGTTAGTAT TAAGAACGTT ATTTATATTT CAAATTTTTC TTTTTTTTCT
     6361 GTACAGACGC GTGTACGCAT GTAACATTAT ACTGAAAACC TTGCTTGAGA AGGTTTTGGG
     6421 ACGCTCGAAG GCTTTAATTT GCAAGCTGGC GCTAATTCAT GGACATGGAG GCCCAGAATA
     6481 CCCTCCTTGA CAGTCTTGAC GTGCGCAGCT CAGGGGCATG ATGTGACTGT CGCCCGTACA
     6541 TTTAGCCCAT ACATCCCCAT GTATAATCAT TTGCATCCAT ACATTTTGAT GGCCGCACGG
     6601 CGCGAAGCAA AAATTACGGC TCCTCGCTGC AGACGTGCGA GCAGGGAAAC GCTCCCCTCA
     6661 CAGACGCGTT GAATTGTCCC CACGCCGCGC CCCTGTAGAG AAATATAAAA GGTTAGGATT
     6721 TGCCACTGAG GTTCTTCTTT CATATACTTC CTTTTAAAAT CTTGCTAGGA TACAGTTCTC
     6781 ACATCACATC CGAACATAAA CAACCATGGG TAAGGAAAAG ACTCACGTTT CGAGGCCGCG
     6841 ATTAAATTCC AACATGGATG CTGATTTATA TGGGTATAAA TGGGCTCGCG ATAATGTCGG
     6901 GCAATCAGGT GCGACAATCT ATCGATTGTA TGGGAAGCCC GATGCGCCAG AGTTGTTTCT
     6961 GAAACATGGC AAAGGTAGCG TTGCCAATGA TGTTACAGAT GAGATGGTCA GACTAAACTG
     7021 GCTGACGGAA TTTATGCCTC TTCCGACCAT CAAGCATTTT ATCCGTACTC CTGATGATGC
     7081 ATGGTTACTC ACCACTGCGA TCCCCGGCAA AACAGCATTC CAGGTATTAG AAGAATATCC
     7141 TGATTCAGGT GAAAATATTG TTGATGCGCT GGCAGTGTTC CTGCGCCGGT TGCATTCGAT
     7201 TCCTGTTTGT AATTGTCCTT TTAACAGCGA TCGCGTATTT CGTCTCGCTC AGGCGCAATC
     7261 ACGAATGAAT AACGGTTTGG TTGATGCGAG TGATTTTGAT GACGAGCGTA ATGGCTGGCC
     7321 TGTTGAACAA GTCTGGAAAG AAATGCATAA GCTTTTGCCA TTCTCACCGG ATTCAGTCGT
     7381 CACTCATGGT GATTTCTCAC TTGATAACCT TATTTTTGAC GAGGGGAAAT TAATAGGTTG
     7441 TATTGATGTT GGACGAGTCG GAATCGCAGA CCGATACCAG GATCTTGCCA TCCTATGGAA
     7501 CTGCCTCGGT GAGTTTTCTC CTTCATTACA GAAACGGCTT TTTCAAAAAT ATGGTATTGA
     7561 TAATCCTGAT ATGAATAAAT TGCAGTTTCA TTTGATGCTC GATGAGTTTT TCTAATCAGT
     7621 ACTGACAATA AAAAGATTCT TGTTTTCAAG AACTTGTCAT TTGTATAGTT TTTTTATATT
     7681 GTAGTTGTTC TATTTTAATC AAATGTTAGC GTGATTTATA TTTTTTTTCG CCTCGACATC
     7741 ATCTGCCCAG ATGCGAAGTT AAGTGCGCAG AAAGTAATAT CATGCGTCAA TCGTATGTGA
     7801 ATGCTGGTCG CTATACTGGC TTCCTTTTCA TCACGTGCTA TAAAAATAAT TATAATTTAA
     7861 ATTTTTTAAT ATAAATATAT AAATTAAAAA TAGAAAGTAA AAAAAGAAAT TAAAGAAAAA
     7921 ATAGTTTTTG TTTTCCGAAG ATGTAAAAGA CTCTAGGGGG ATCGCCAACA AATACTACCT
     7981 TTTATCTTGC TCTTCCTGCT CTCAGGTATT AATGCCGAAT TGTTTCATCT TGTCTGTGTA
     8041 GAACACCACA CACGAAAATC CTGTGATTTT ACATTTTACT TATCGTTAAT CGAATGTATA
     8101 TCTATTTAAT CTGCTTTTCT TGTCTAATAA ATATATATGT AAAGTACGCT TTTTGTTGAA
     8161 ATTTTTTAAA CCTTTGTTTA TTTTTTTTTC TTCATTCCGT AACTCTTCTA CCTTCTTTAT
     8221 TTACTTTCTA AAATCCAAAT ACAAAACATA AAAATAAATA AACACAGAGT AAATTCCCAA
     8281 ATTATTCCAT CATTAAAAGA TACGAGGCGC GTGTAAGTTA CAGGCA
//
LOCUS       BB3_pGAP_Linker_BbsI(intern)_RPS25Att_pLAT1_Cas9_cycTTG_418      9103 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Vector          join(7231..9103,1..730)
                     /origin="BB3_pTEF_KanR_TEFt_AC (2,633bp circular)Target
                     vector: from BsaI cut at 760^761 to BsaI cut at 730^731,
                     2,603bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     rep_origin      complement(13..686)
                     /label="pUC_ORI_aus_pGAPZB"
     primer          524..541
                     /label="B1_13"
     terminator      691..730
                     /label="BBa_B1007"
     FS              731..734
                     /label="FS A"
     Vector          join(1751..>1754,<731..734)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 734^735
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        737..>1229
                     /label="pGAP"
     Restriction_sit join(1229^1230,complement(1229^1230))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1230..>1261
                     /label="L_FS23_BbsI_fw"
     Linker          1230..1261
                     /origin="PCR_Linker_FS23_BbsI(intern) (54bp
                     linear)Fragment: from BpiI cut at 9^10 to BpiI cut at
                     41^42, 32bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="FS 23 Linker"
     FS              1230..1233
                     /label="FS2"
     primer          complement(<1234..>1265)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1262..1294,complement(1295..1727),complement(1728..>1754))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1266..>1750
                     /note="Forward primer name: RPS25ATT_M1_FW"
                     /note="Forward primer: GacagGtcTccgCTTCCTTGTCGAGTGTGTTT"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_RV"
                     /note="Reverse primer:
                     gatcggtctCcAgctGTCAAATATTCAGATGGTGA"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 458"
                     /label="rps25aTT"
     FS              1751..1754
                     /label="FS 4"
     primer          <1755..>1758
                     /label="3_BsaI_FsB_Bam"
     misc_feature    join(<7227..>7230,<1755..>1758)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (108bp linear)Fragment: full
                     sequenceOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="Insert"
     FS              1755..1758
                     /label="FS B"
     Vector          join(7227..>7230,<1755..1758)
                     /origin="BB2_L_BC_syn_BbsI_7 (2,215bp circular)Target
                     vector: from BpiI cut at 639^640 to BpiI cut at 607^608,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          complement(<1759..>1762)
                     /label="4_BsaI_FsB_BamXho"
     promoter        <1759..>2763
                     /locus_tag="PAS_chr1-1_0049"
                     /codon_start=1
                     /product="Ribonuclease H2 catalytic subunit, removes RNA
                     primers during Okazaki fragment synthesis"
                     /protein_id="XP_002489688.1"
                     /db_xref="GI:254565155"
                     /db_xref="GeneID:8197566"
                     /translation="MTRIENRVCDSSPMTTTFIPPSVPEISTYRTTTYVSEIPESVLK
                     NPEKPCCLGVDEAGRGPVVGPMVYGVSYCLEEYEPRLKDFGFADSKTLTEQKRSELLE
                     SLCNGDHELRKNVGWATTSISACDISSAMLKPHNIGNYNLNEQAHDVTMELIDKVIQK
                     GVNLRHVYVDTVGPPASYQKKLQQRFPALLITVSKKADAIYPCVSSASVCAKVTRDVC
                     LRTSNGEENLLDCGSGYPSDPNTKKWLAQSIHPLFGWSRDVRFSWQTAKDCLERHNSI
                     KVVFGEEEPTPVNFGDLGLSKQKTDFKDLEISTSWWGH"
                     /label="pLAT1"
     Site            join(2763^2764,complement(2763^2764),complement(2763^2764))
                     /site_type="restriction site"
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 3162^3163"
                     /note="cut_0_on_negative_strand: 3166^3167"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <2764..2788
                     /label="FS2_Cas9_BsaI_fw"
     CDS             2765..6904
                     /label="Cas9"
     Taming          2815
                     /label="C>T"
     primer          complement(4176..4195)
                     /label="1_cas9_rev"
     primer          4176..4195
                     /label="1_cas9_fw"
     Taming          4180
                     /label="C>t"
     primer          complement(4450..4471)
                     /label="2_cas9_rev"
     primer          4450..4471
                     /label="2_cas9_fw"
     Taming          4462
                     /label="A>g"
     primer          complement(4919..4936)
                     /label="3_cas9_rev"
     primer          4919..4936
                     /label="3_cas9_fw"
     Taming          4921
                     /label="T>c"
     primer          complement(5634..5655)
                     /label="4_cas9_rev"
     primer          5634..5655
                     /label="4_cas9_fw"
     Taming          5650
                     /label="G>c"
     primer          complement(6501..6518)
                     /label="5_cas9_rev"
     primer          6501..6519
                     /label="5_cas9_fw"
     Taming          6508
                     /label="T>c"
     NLS             6869..6904
                     /label="NLS SV40"
     primer          complement(6882..>6908)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6905..>7226
                     /label="FS3_CYCtt_FS4"
     primer          <7227..>7230
                     /label="5_BsaI_FsC_Xho"
     FS              7227..7230
                     /label="FS 4"
     primer          complement(7231..>7234)
                     /label="6_BsaI_FsC"
     promoter        7239..7582
                     /label="pTEF (Ashbya gossypii)"
     mutation        7412
                     /label="C to G"
     CDS             7583..8392
                     /label="KanR / G418"
     terminator      8401..8599
                     /label="TEFtt (Ashbya gossypii)"
     misc_feature    8600..9103
                     /label="CEN6_ARS4"
     mutation        8821
                     /label="G/A, actually G/C"
ORIGIN
        1 AGCGATCGCG CTGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT
       61 GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG
      121 TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC
      181 CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC
      241 TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT
      301 CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT
      361 ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC
      421 AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA
      481 GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG CTCTGCTGAA
      541 GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG
      601 TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA
      661 AGATCCTTTG ATCTTTTCTA CGGGGTGGAG cgcaaaaaac cccgcccctg acagggcggg
      721 gttttttcgc GATCGGAGGA TCCTTTTTTG TAGAAATGTC TTGGTGTCCT CGTCCAATCA
      781 GGTAGCCATC TCTGAAATAT CTGGCTCCGT TGCAACTCCG AACGACCTGC TGGCAACGTA
      841 AAATTCTCCG GGGTAAAACT TAAATGTGGA GTAATGGAAC CAGAAACGTC TCTTCCCTTC
      901 TCTCTCCTTC CACCGCCCGT TACCGTCCCT AGGAAATTTT ACTCTGCTGG AGAGCTTCTT
      961 CTACGGCCCC CTTGCAGCAA TGCTCTTCCC AGCATTACGT TGCGGGTAAA ACGGAGGTCG
     1021 TGTACCCGAC CTAGCAGCCC AGGGATGGAA AAGTCCCGGC CGTCGCTGGC AATAATAGCG
     1081 GGCGGACGCA TGTCATGAGA TTATTGGAAA CCACCAGAAT CGAATATAAA AGGCGAACAC
     1141 CTTTCCCAAT TTTGGTTTCT CCTGACCCAA AGACTTTAAA TTTAATTTAT TTGTCCCTAT
     1201 TTCAATCAAT TGAACAACTA TCAAAACACC ATGGCGTCTT CGGATCCCTC GAGGAAGACG
     1261 CGCTTTTAGT GTACATCTGA TAATATAGTA CTACCACGTA TGATAATGTA GAGAATAGGC
     1321 TTCCTTGTCG AGTGTGTTTG CAGTTTTCTT GAGTTTCAAG GTTTAAATGC TGGTATATTA
     1381 GTTCATCGAA GGTTTCAGCC AATAGCACCT TAAATCAATC AAACTAATTC GACTCTTACG
     1441 AAAGAGCCTA CTGTGTTTAG TATCGAAGTC GTTTACCTTT CATGTTGAAT AGCTTCCTCT
     1501 CTGACCCTAA CATTTCAAGA TCCTCCTAAA GTTACCCGGA TTGTGAAATT CTAATGATCC
     1561 ACCTGCCCAA TGCATTTTTT CTTTATTCAG TTTACCTTTT TTACCTAATA TACGAGCTTG
     1621 TTAAAGTAAG TGGCACTGCA ATACTAGGCT TATTGTTGAT ATTATGATGA ATCGTTTTCA
     1681 CAAACTTGAT TTCCTGTGAA CTCACCATGT ACTAAGGAAA AAAACATGCA TCACCATCTG
     1741 AATATTTGAC CGCTCCGGGG AGGGAAGAAT ATGAACCACG ATTGAAAGAC TTTGGGTTTG
     1801 CCGATTCTAA AACACTTACT GAACAGAAAA GAAGTGAATT ATTAGAGAGT TTATGCAACG
     1861 GAGATCATGA GCTTAGAAAG AACGTAGGCT GGGCAACAAC ATCCATTTCT GCCTGCGATA
     1921 TTTCCAGTGC AATGCTAAAG CCACATAATA TAGGCAATTA CAACTTGAAT GAGCAGGCTC
     1981 ATGATGTCAC CATGGAACTG ATAGATAAAG TCATCCAAAA AGGTGTCAAT TTACGGCACG
     2041 TCTACGTAGA TACTGTCGGA CCTCCAGCAT CATACCAAAA GAAACTGCAA CAAAGGTTTC
     2101 CCGCACTGCT GATCACCGTA TCTAAAAAAG CAGATGCCAT CTATCCATGT GTTTCAAGTG
     2161 CTTCAGTTTG CGCCAAGGTC ACTAGAGACG TCTGTTTGAG AACCTCAAAT GGGGAGGAGA
     2221 ACCTATTAGA TTGTGGGTCT GGATATCCCT CGGACCCAAA TACTAAGAAA TGGCTAGCCC
     2281 AGTCTATACA TCCATTGTTT GGCTGGAGTC GTGATGTCAG GTTTTCGTGG CAGACGGCCA
     2341 AAGATTGTCT TGAGAGACAC AACTCAATCA AAGTTGTGTT TGGCGAAGAG GAGCCTACCC
     2401 CGGTTAATTT CGGTGATCTG GGGTTGAGCA AACAGAAGCC CGACTTTAAG GATTTAGAAA
     2461 TCAGTACCAG TTGGTGGGGA CATTAATTGG TGGAGGAGAG CCAATATTCT CAACTGTTCG
     2521 TAGGACCATA ACCCCCCACT CGCACCAGAA ATACTCATTG GCGCTGACAA CTAGTTTGAG
     2581 CCGCTTCAAT TGCGGCAAGT CCCCAGTTTT GTTTGGTACT CCCCTCTCGC CTCGCGTGTT
     2641 TTTGTTCCTC CGAATTGTGC ATCACCCTGT CACATCCGTA CACCATCGGA ACACTCTGCT
     2701 CCTCCTCCTT TCTCCTCTTA ATTGAACGTA TATCTAATTA CCCTAATAAC AAGTATCTTC
     2761 AACCATGGAC AAGAAGTACT CCATTGGGCT CGATATCGGC ACAAACAGCG TCGGTTGGGC
     2821 CGTCATTACG GACGAGTACA AGGTGCCGAG CAAAAAATTC AAAGTTCTGG GCAATACCGA
     2881 TCGCCACAGC ATAAAGAAGA ACCTCATTGG CGCCCTCCTG TTCGACTCCG GGGAGACGGC
     2941 CGAAGCCACG CGGCTCAAAA GAACAGCACG GCGCAGATAT ACCCGCAGAA AGAATCGGAT
     3001 CTGCTACCTG CAGGAGATCT TTAGTAATGA GATGGCTAAG GTGGATGACT CTTTCTTCCA
     3061 TAGGCTGGAG GAGTCCTTTT TGGTGGAGGA GGATAAAAAG CACGAGCGCC ACCCAATCTT
     3121 TGGCAATATC GTGGACGAGG TGGCGTACCA TGAAAAGTAC CCAACCATAT ATCATCTGAG
     3181 GAAGAAGCTT GTAGACAGTA CTGATAAGGC TGACTTGCGG TTGATCTATC TCGCGCTGGC
     3241 GCATATGATC AAATTTCGGG GACACTTCCT CATCGAGGGG GACCTGAACC CAGACAACAG
     3301 CGATGTCGAC AAACTCTTTA TCCAACTGGT TCAGACTTAC AATCAGCTTT TCGAAGAGAA
     3361 CCCGATCAAC GCATCCGGAG TTGACGCCAA AGCAATCCTG AGCGCTAGGC TGTCCAAATC
     3421 CCGGCGGCTC GAAAACCTCA TCGCACAGCT CCCTGGGGAG AAGAAGAACG GCCTGTTTGG
     3481 TAATCTTATC GCCCTGTCAC TCGGGCTGAC CCCCAACTTT AAATCTAACT TCGACCTGGC
     3541 CGAAGATGCC AAGCTTCAAC TGAGCAAAGA CACCTACGAT GATGATCTCG ACAATCTGCT
     3601 GGCCCAGATC GGCGACCAGT ACGCAGACCT TTTTTTGGCG GCAAAGAACC TGTCAGACGC
     3661 CATTCTGCTG AGTGATATTC TGCGAGTGAA CACGGAGATC ACCAAAGCTC CGCTGAGCGC
     3721 TAGTATGATC AAGCGCTATG ATGAGCACCA CCAAGACTTG ACTTTGCTGA AGGCCCTTGT
     3781 CAGACAGCAA CTGCCTGAGA AGTACAAGGA AATTTTCTTC GATCAGTCTA AAAATGGCTA
     3841 CGCCGGATAC ATTGACGGCG GAGCAAGCCA GGAGGAATTT TACAAATTTA TTAAGCCCAT
     3901 CTTGGAAAAA ATGGACGGCA CCGAGGAGCT GCTGGTAAAG CTTAACAGAG AAGATCTGTT
     3961 GCGCAAACAG CGCACTTTCG ACAATGGAAG CATCCCCCAC CAGATTCACC TGGGCGAACT
     4021 GCACGCTATC CTCAGGCGGC AAGAGGATTT CTACCCCTTT TTGAAAGATA ACAGGGAAAA
     4081 GATTGAGAAA ATCCTCACAT TTCGGATACC CTACTATGTA GGCCCCCTCG CCCGGGGAAA
     4141 TTCCAGATTC GCGTGGATGA CTCGCAAATC AGAAGAGACt ATCACTCCCT GGAACTTCGA
     4201 GGAAGTCGTG GATAAGGGGG CCTCTGCCCA GTCCTTCATC GAAAGGATGA CTAACTTTGA
     4261 TAAAAATCTG CCTAACGAAA AGGTGCTTCC TAAACACTCT CTGCTGTACG AGTACTTCAC
     4321 AGTTTATAAC GAGCTCACCA AGGTCAAATA CGTCACAGAA GGGATGAGAA AGCCAGCATT
     4381 CCTGTCTGGA GAGCAGAAGA AAGCTATCGT GGACCTCCTC TTCAAGACGA ACCGGAAAGT
     4441 TACCGTGAAA CAGCTCAAAG AgGACTATTT CAAAAAGATT GAATGTTTCG ACTCTGTTGA
     4501 AATCAGCGGA GTGGAGGATC GCTTCAACGC ATCCCTGGGA ACGTATCACG ATCTCCTGAA
     4561 AATCATTAAA GACAAGGACT TCCTGGACAA TGAGGAGAAC GAGGACATTC TTGAGGACAT
     4621 TGTCCTCACC CTTACGTTGT TTGAAGATAG GGAGATGATT GAAGAACGCT TGAAAACTTA
     4681 CGCTCATCTC TTCGACGACA AAGTCATGAA ACAGCTCAAG AGGCGCCGAT ATACAGGATG
     4741 GGGGCGGCTG TCAAGAAAAC TGATCAATGG GATCCGAGAC AAGCAGAGTG GAAAGACAAT
     4801 CCTGGATTTT CTTAAGTCCG ATGGATTTGC CAACCGGAAC TTCATGCAGT TGATCCATGA
     4861 TGACTCTCTC ACCTTTAAGG AGGACATCCA GAAAGCACAA GTTTCTGGCC AGGGGGACAG
     4921 cCTTCACGAG CACATCGCTA ATCTTGCAGG TAGCCCAGCT ATCAAAAAGG GAATACTGCA
     4981 GACCGTTAAG GTCGTGGATG AACTCGTCAA AGTAATGGGA AGGCATAAGC CCGAGAATAT
     5041 CGTTATCGAG ATGGCCCGAG AGAACCAAAC TACCCAGAAG GGACAGAAGA ACAGTAGGGA
     5101 AAGGATGAAG AGGATTGAAG AGGGTATAAA AGAACTGGGG TCCCAAATCC TTAAGGAACA
     5161 CCCAGTTGAA AACACCCAGC TTCAGAATGA GAAGCTCTAC CTGTACTACC TGCAGAACGG
     5221 CAGGGACATG TACGTGGATC AGGAACTGGA CATCAATCGG CTCTCCGACT ACGACGTGGA
     5281 TCATATCGTG CCCCAGTCTT TTCTCAAAGA TGATTCTATT GATAATAAAG TGTTGACAAG
     5341 ATCCGATAAA AATAGAGGGA AGAGTGATAA CGTCCCCTCA GAAGAAGTTG TCAAGAAAAT
     5401 GAAAAATTAT TGGCGGCAGC TGCTGAACGC CAAACTGATC ACACAACGGA AGTTCGATAA
     5461 TCTGACTAAG GCTGAACGAG GTGGCCTGTC TGAGTTGGAT AAAGCCGGCT TCATCAAAAG
     5521 GCAGCTTGTT GAGACACGCC AGATCACCAA GCACGTGGCC CAAATTCTCG ATTCACGCAT
     5581 GAACACCAAG TACGATGAAA ATGACAAACT GATTCGAGAG GTGAAAGTTA TTACTCTGAA
     5641 GTCTAAGCTc GTCTCAGATT TCAGAAAGGA CTTTCAGTTT TATAAGGTGA GAGAGATCAA
     5701 CAATTACCAC CATGCGCATG ATGCCTACCT GAATGCAGTG GTAGGCACTG CACTTATCAA
     5761 AAAATATCCC AAGCTTGAAT CTGAATTTGT TTACGGAGAC TATAAAGTGT ACGATGTTAG
     5821 GAAAATGATC GCAAAGTCTG AGCAGGAAAT AGGCAAGGCC ACCGCTAAGT ACTTCTTTTA
     5881 CAGCAATATT ATGAATTTTT TCAAGACCGA GATTACACTG GCCAATGGAG AGATTCGGAA
     5941 GCGACCACTT ATCGAAACAA ACGGAGAAAC AGGAGAAATC GTGTGGGACA AGGGTAGGGA
     6001 TTTCGCGACA GTCCGGAAGG TCCTGTCCAT GCCGCAGGTG AACATCGTTA AAAAGACCGA
     6061 AGTACAGACC GGAGGCTTCT CCAAGGAAAG TATCCTCCCG AAAAGGAACA GCGACAAGCT
     6121 GATCGCACGC AAAAAAGATT GGGACCCCAA GAAATACGGC GGATTCGATT CTCCTACAGT
     6181 CGCTTACAGT GTACTGGTTG TGGCCAAAGT GGAGAAAGGG AAGTCTAAAA AACTCAAAAG
     6241 CGTCAAGGAA CTGCTGGGCA TCACAATCAT GGAGCGATCA AGCTTCGAAA AAAACCCCAT
     6301 CGACTTTCTC GAGGCGAAAG GATATAAAGA GGTCAAAAAA GACCTCATCA TTAAGCTTCC
     6361 CAAGTACTCT CTCTTTGAGC TTGAAAACGG CCGGAAACGA ATGCTCGCTA GTGCGGGCGA
     6421 GCTGCAGAAA GGTAACGAGC TGGCACTGCC CTCTAAATAC GTTAATTTCT TGTATCTGGC
     6481 CAGCCACTAT GAAAAGCTCA AAGGGTCcCC CGAAGATAAT GAGCAGAAGC AGCTGTTCGT
     6541 GGAACAACAC AAACACTACC TTGATGAGAT CATCGAGCAA ATAAGCGAAT TCTCCAAAAG
     6601 AGTGATCCTC GCCGACGCTA ACCTCGATAA GGTGCTTTCT GCTTACAATA AGCACAGGGA
     6661 TAAGCCCATC AGGGAGCAGG CAGAAAACAT TATCCACTTG TTTACTCTGA CCAACTTGGG
     6721 CGCGCCTGCA GCCTTCAAGT ACTTCGACAC CACCATAGAC AGAAAGCGGT ACACCTCTAC
     6781 AAAGGAGGTC CTGGACGCCA CACTGATTCA TCAGTCAATT ACGGGGCTCT ATGAAACAAG
     6841 AATCGACCTC TCTCAGCTCG GTGGAGACAG CAGGGCTGAC CCCAAGAAGA AGAGGAAGGT
     6901 GTGAGCTTCG TCCGACGGCG GCCCACGGGT CCCAGGCCTC GGAGATCCGT CCCCCTTTTC
     6961 CTTTGTCGAT ATCATGTAAT TAGTTATGTC ACGCTTACAT TCACGCCCTC CCCCCACATC
     7021 CGCTCTAACC GAAAAGGAAG GAGTTAGACA ACCTGAAGTC TAGGTCCCTA TTTATTTTTT
     7081 TATAGTTATG TTAGTATTAA GAACGTTATT TATATTTCAA ATTTTTCTTT TTTTTCTGTA
     7141 CAGACGCGTG TACGCATGTA ACATTATACT GAAAACCTTG CTTGAGAAGG TTTTGGGACG
     7201 CTCGAAGGCT TTAATTTGCA AGCTGGCGCT AATTCATGGA CATGGAGGCC CAGAATACCC
     7261 TCCTTGACAG TCTTGACGTG CGCAGCTCAG GGGCATGATG TGACTGTCGC CCGTACATTT
     7321 AGCCCATACA TCCCCATGTA TAATCATTTG CATCCATACA TTTTGATGGC CGCACGGCGC
     7381 GAAGCAAAAA TTACGGCTCC TCGCTGCAGA CGTGCGAGCA GGGAAACGCT CCCCTCACAG
     7441 ACGCGTTGAA TTGTCCCCAC GCCGCGCCCC TGTAGAGAAA TATAAAAGGT TAGGATTTGC
     7501 CACTGAGGTT CTTCTTTCAT ATACTTCCTT TTAAAATCTT GCTAGGATAC AGTTCTCACA
     7561 TCACATCCGA ACATAAACAA CCATGGGTAA GGAAAAGACT CACGTTTCGA GGCCGCGATT
     7621 AAATTCCAAC ATGGATGCTG ATTTATATGG GTATAAATGG GCTCGCGATA ATGTCGGGCA
     7681 ATCAGGTGCG ACAATCTATC GATTGTATGG GAAGCCCGAT GCGCCAGAGT TGTTTCTGAA
     7741 ACATGGCAAA GGTAGCGTTG CCAATGATGT TACAGATGAG ATGGTCAGAC TAAACTGGCT
     7801 GACGGAATTT ATGCCTCTTC CGACCATCAA GCATTTTATC CGTACTCCTG ATGATGCATG
     7861 GTTACTCACC ACTGCGATCC CCGGCAAAAC AGCATTCCAG GTATTAGAAG AATATCCTGA
     7921 TTCAGGTGAA AATATTGTTG ATGCGCTGGC AGTGTTCCTG CGCCGGTTGC ATTCGATTCC
     7981 TGTTTGTAAT TGTCCTTTTA ACAGCGATCG CGTATTTCGT CTCGCTCAGG CGCAATCACG
     8041 AATGAATAAC GGTTTGGTTG ATGCGAGTGA TTTTGATGAC GAGCGTAATG GCTGGCCTGT
     8101 TGAACAAGTC TGGAAAGAAA TGCATAAGCT TTTGCCATTC TCACCGGATT CAGTCGTCAC
     8161 TCATGGTGAT TTCTCACTTG ATAACCTTAT TTTTGACGAG GGGAAATTAA TAGGTTGTAT
     8221 TGATGTTGGA CGAGTCGGAA TCGCAGACCG ATACCAGGAT CTTGCCATCC TATGGAACTG
     8281 CCTCGGTGAG TTTTCTCCTT CATTACAGAA ACGGCTTTTT CAAAAATATG GTATTGATAA
     8341 TCCTGATATG AATAAATTGC AGTTTCATTT GATGCTCGAT GAGTTTTTCT AATCAGTACT
     8401 GACAATAAAA AGATTCTTGT TTTCAAGAAC TTGTCATTTG TATAGTTTTT TTATATTGTA
     8461 GTTGTTCTAT TTTAATCAAA TGTTAGCGTG ATTTATATTT TTTTTCGCCT CGACATCATC
     8521 TGCCCAGATG CGAAGTTAAG TGCGCAGAAA GTAATATCAT GCGTCAATCG TATGTGAATG
     8581 CTGGTCGCTA TACTGGCTTC CTTTTCATCA CGTGCTATAA AAATAATTAT AATTTAAATT
     8641 TTTTAATATA AATATATAAA TTAAAAATAG AAAGTAAAAA AAGAAATTAA AGAAAAAATA
     8701 GTTTTTGTTT TCCGAAGATG TAAAAGACTC TAGGGGGATC GCCAACAAAT ACTACCTTTT
     8761 ATCTTGCTCT TCCTGCTCTC AGGTATTAAT GCCGAATTGT TTCATCTTGT CTGTGTAGAA
     8821 CACCACACAC GAAAATCCTG TGATTTTACA TTTTACTTAT CGTTAATCGA ATGTATATCT
     8881 ATTTAATCTG CTTTTCTTGT CTAATAAATA TATATGTAAA GTACGCTTTT TGTTGAAATT
     8941 TTTTAAACCT TTGTTTATTT TTTTTTCTTC ATTCCGTAAC TCTTCTACCT TCTTTATTTA
     9001 CTTTCTAAAA TCCAAATACA AAACATAAAA ATAAATAAAC ACAGAGTAAA TTCCCAAATT
     9061 ATTCCATCAT TAAAAGATAC GAGGCGCGTG TAAGTTACAG GCA
//
LOCUS       BB3_pGAP_BbsI(intern)_RPS25Att_pTEF_Cas9_cycTT_G418      8513 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     Vector          join(6641..8513,1..730)
                     /origin="BB3_pTEF_KanR_TEFt_AC (2,633bp circular)Target
                     vector: from BsaI cut at 760^761 to BsaI cut at 730^731,
                     2,603bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     rep_origin      complement(13..686)
                     /label="pUC_ORI_aus_pGAPZB"
     primer          448..467
                     /label="JH_77_fw"
     primer          524..541
                     /label="B1_13"
     terminator      691..730
                     /label="BBa_B1007"
     FS              731..734
                     /label="FS A"
     Vector          join(1751..>1754,<731..734)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 796^797 to BpiI cut at 764^765,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Restriction_sit 734^735
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2153^2154"
                     /note="cut_0_on_negative_strand: 2157^2158"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     promoter        737..>1229
                     /label="pGAP"
     Restriction_sit join(1229^1230,complement(1229^1230))
                     /note="Name: BbsI"
                     /note="Pattern: gaagac"
                     /note="Number_of_matches: 2"
                     /note="cut_0_on_positive_strand: 2648^2649"
                     /note="cut_0_on_negative_strand: 2652^2653"
                     /note="inhibited_by: "
                     /note="site_type: other"
                     /note="restriction site"
                     /label="BbsI"
     primer          <1230..>1261
                     /label="L_FS23_BbsI_fw"
     Linker          1230..1265
                     /label="FS 23 Linker"
     FS              1230..1233
                     /label="FS2"
     primer          complement(<1234..>1265)
                     /label="L_FS23_BbsI_rev"
     misc_feature    join(<1262..1294,complement(1295..1727),complement(1728..>1754))
                     /feature_type="PCR fragment"
                     /note="Forward primer name: RPS25ATT_rv_fs4"
                     /note="Forward primer:
                     gAtcgGtctCcAGCgGTCAAATATTCAGATGGTGATGC"
                     /note="Forward primer misprimes: 9"
                     /note="Reverse primer name: RPS25ATT_FW_fs3"
                     /note="Reverse primer:
                     gaCaggtctcCgcttTTAGTGTACATCTGATAATATAGTACTAC"
                     /note="Reverse primer misprimes: 13"
                     /note="Fragment length: 515"
                     /label="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3))"
     terminator      1262..1750
                     /origin="Fragment (TT_BB1_ RPS25Att (RPS25ATT_rv_fs4 -
                     RPS25ATT_FW_fs3)) (515bp linear)Fragment: from BsaI cut
                     at 11^12 to BsaI cut at 500^501, 489bpOrientation:
                     reverse complementedPlus strand 5':   no changePlus
                     strand 3':   no changeMinus strand 5':  no changeMinus
                     strand 3':  no change"
                     /label="RPS25Att"
     FS              1751..1754
                     /label="FS 4"
     misc_feature    join(<6637..>6640,<1755..>1758)
                     /feature_type="Insertion"
                     /origin="BsaI_B_1_4_C_FW (118bp linear)Fragment: from
                     SpeI cut at 5^6 to AgeI cut at 109^110, 104bpOrientation:
                     originalPlus strand 5':   no changePlus strand 3':   no
                     changeMinus strand 5':  no changeMinus strand 3':  no
                     change"
                     /label="Insert"
     Vector          join(6637..>6640,<1755..1758)
                     /origin="PCRproductNewBB2_BsaI_B_1_4_C_FW (2.215bp
                     circular)Target vector: from BbsI cut at 2199^2200 to
                     BbsI cut at 2167^2168, 2.183bpOrientation: originalPlus
                     strand 5':   no changePlus strand 3':   no changeMinus
                     strand 5':  no changeMinus strand 3':  no change"
     promoter        1763..2173
                     /label="TEF1p"
     primer          <2174..2198
                     /label="FS2_Cas9_BsaI_fw"
     CDS             2175..6314
                     /label="Cas9"
     Taming          2225
                     /label="C>T"
     primer          complement(3586..3605)
                     /label="1_cas9_rev"
     primer          3586..3605
                     /label="1_cas9_fw"
     Taming          3590
                     /label="C>t"
     primer          complement(3860..3881)
                     /label="2_cas9_rev"
     primer          3860..3881
                     /label="2_cas9_fw"
     Taming          3872
                     /label="A>g"
     primer          complement(4329..4346)
                     /label="3_cas9_rev"
     primer          4329..4346
                     /label="3_cas9_fw"
     Taming          4331
                     /label="T>c"
     primer          complement(5044..5065)
                     /label="4_cas9_rev"
     primer          5044..5065
                     /label="4_cas9_fw"
     Taming          5060
                     /label="G>c"
     primer          complement(5911..5928)
                     /label="5_cas9_rev"
     primer          5911..5929
                     /label="5_cas9_fw"
     Taming          5918
                     /label="T>c"
     NLS             6279..6314
                     /label="NLS SV40"
     primer          complement(6292..>6318)
                     /label="FS3_Pol7_Cas9_BsaI_rev"
     terminator      <6315..>6636
                     /label="FS3_CYCtt_FS4"
     primer          complement(6441..6460)
                     /label="Seq cycTT Rev1"
     CDS             6649..6992
                     /label="TEFpromoter"
     mutation        6822
                     /label="C to G"
     primer          6851..6868
                     /label="Seq_MX_FW"
     primer          complement(6876..6895)
                     /label="JH_78_rev"
     CDS             6993..7802
                     /label="KanR"
     CDS             7811..8009
                     /label="TEFterminator"
     primer          complement(7834..7855)
                     /label="Seq_MX_REV"
     misc_feature    8010..8513
                     /label="CEN6_ARS4"
     mutation        8231
                     /label="G/A, actually G/C"
ORIGIN
        1 AGCGATCGCG CTGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT
       61 GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG
      121 TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC
      181 CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC
      241 TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT
      301 CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT
      361 ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC
      421 AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA
      481 GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG CTCTGCTGAA
      541 GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG
      601 TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA
      661 AGATCCTTTG ATCTTTTCTA CGGGGTGGAG cgcaaaaaac cccgcccctg acagggcggg
      721 gttttttcgc GATCGGAGGA TCCTTTTTTG TAGAAATGTC TTGGTGTCCT CGTCCAATCA
      781 GGTAGCCATC TCTGAAATAT CTGGCTCCGT TGCAACTCCG AACGACCTGC TGGCAACGTA
      841 AAATTCTCCG GGGTAAAACT TAAATGTGGA GTAATGGAAC CAGAAACGTC TCTTCCCTTC
      901 TCTCTCCTTC CACCGCCCGT TACCGTCCCT AGGAAATTTT ACTCTGCTGG AGAGCTTCTT
      961 CTACGGCCCC CTTGCAGCAA TGCTCTTCCC AGCATTACGT TGCGGGTAAA ACGGAGGTCG
     1021 TGTACCCGAC CTAGCAGCCC AGGGATGGAA AAGTCCCGGC CGTCGCTGGC AATAATAGCG
     1081 GGCGGACGCA TGTCATGAGA TTATTGGAAA CCACCAGAAT CGAATATAAA AGGCGAACAC
     1141 CTTTCCCAAT TTTGGTTTCT CCTGACCCAA AGACTTTAAA TTTAATTTAT TTGTCCCTAT
     1201 TTCAATCAAT TGAACAACTA TCAAAACACC ATGGCGTCTT CGGATCCCTC GAGGAAGACG
     1261 CGCTTTTAGT GTACATCTGA TAATATAGTA CTACCACGTA TGATAATGTA GAGAATAGGC
     1321 TTCCTTGTCG AGTGTGTTTG CAGTTTTCTT GAGTTTCAAG GTTTAAATGC TGGTATATTA
     1381 GTTCATCGAA GGTTTCAGCC AATAGCACCT TAAATCAATC AAACTAATTC GACTCTTACG
     1441 AAAGAGCCTA CTGTGTTTAG TATCGAAGTC GTTTACCTTT CATGTTGAAT AGCTTCCTCT
     1501 CTGACCCTAA CATTTCAAGA TCCTCCTAAA GTTACCCGGA TTGTGAAATT CTAATGATCC
     1561 ACCTGCCCAA TGCATTTTTT CTTTATTCAG TTTACCTTTT TTACCTAATA TACGAGCTTG
     1621 TTAAAGTAAG TGGCACTGCA ATACTAGGCT TATTGTTGAT ATTATGATGA ATCGTTTTCA
     1681 CAAACTTGAT TTCCTGTGAA CTCACCATGT ACTAAGGAAA AAAACATGCA TCACCATCTG
     1741 AATATTTGAC CGCTCCGGGG AGCACACACC ATAGCTTCAA AATGTTTCTA CTCCTTTTTT
     1801 ACTCTTCCAG ATTTTCTCGG ACTCCGCGCA TCGCCGTACC ACTTCAAAAC ACCCAAGCAC
     1861 AGCATACTAA ATTTTCCCTC TTTCTTCCTC TAGGGTGTCG TTAATTACCC GTACTAAAGG
     1921 TTTGGAAAAG AAAAAAGcGA CCGCCTCGTT TCTTTTTCTT CGTCGAAAAA GGCAATAAAA
     1981 ATTTTTATCA CGTTTCTTTT TCTTGAAATT TTTTTTTTTA GTTTTTTTCT CTTTCAGTGA
     2041 CCTCCATTGA TATTTAAGTT AATAAACGGT gTTCAATTTC TCAAGTTTCA GTTTCATTTT
     2101 TCTTGTTCTA TTACAACTTT TTTTACTTCT TGTTCATTAG AAAGAAAGCA TAGCAATCTA
     2161 ATCTAAGGGC GGTCATGGAC AAGAAGTACT CCATTGGGCT CGATATCGGC ACAAACAGCG
     2221 TCGGTTGGGC CGTCATTACG GACGAGTACA AGGTGCCGAG CAAAAAATTC AAAGTTCTGG
     2281 GCAATACCGA TCGCCACAGC ATAAAGAAGA ACCTCATTGG CGCCCTCCTG TTCGACTCCG
     2341 GGGAGACGGC CGAAGCCACG CGGCTCAAAA GAACAGCACG GCGCAGATAT ACCCGCAGAA
     2401 AGAATCGGAT CTGCTACCTG CAGGAGATCT TTAGTAATGA GATGGCTAAG GTGGATGACT
     2461 CTTTCTTCCA TAGGCTGGAG GAGTCCTTTT TGGTGGAGGA GGATAAAAAG CACGAGCGCC
     2521 ACCCAATCTT TGGCAATATC GTGGACGAGG TGGCGTACCA TGAAAAGTAC CCAACCATAT
     2581 ATCATCTGAG GAAGAAGCTT GTAGACAGTA CTGATAAGGC TGACTTGCGG TTGATCTATC
     2641 TCGCGCTGGC GCATATGATC AAATTTCGGG GACACTTCCT CATCGAGGGG GACCTGAACC
     2701 CAGACAACAG CGATGTCGAC AAACTCTTTA TCCAACTGGT TCAGACTTAC AATCAGCTTT
     2761 TCGAAGAGAA CCCGATCAAC GCATCCGGAG TTGACGCCAA AGCAATCCTG AGCGCTAGGC
     2821 TGTCCAAATC CCGGCGGCTC GAAAACCTCA TCGCACAGCT CCCTGGGGAG AAGAAGAACG
     2881 GCCTGTTTGG TAATCTTATC GCCCTGTCAC TCGGGCTGAC CCCCAACTTT AAATCTAACT
     2941 TCGACCTGGC CGAAGATGCC AAGCTTCAAC TGAGCAAAGA CACCTACGAT GATGATCTCG
     3001 ACAATCTGCT GGCCCAGATC GGCGACCAGT ACGCAGACCT TTTTTTGGCG GCAAAGAACC
     3061 TGTCAGACGC CATTCTGCTG AGTGATATTC TGCGAGTGAA CACGGAGATC ACCAAAGCTC
     3121 CGCTGAGCGC TAGTATGATC AAGCGCTATG ATGAGCACCA CCAAGACTTG ACTTTGCTGA
     3181 AGGCCCTTGT CAGACAGCAA CTGCCTGAGA AGTACAAGGA AATTTTCTTC GATCAGTCTA
     3241 AAAATGGCTA CGCCGGATAC ATTGACGGCG GAGCAAGCCA GGAGGAATTT TACAAATTTA
     3301 TTAAGCCCAT CTTGGAAAAA ATGGACGGCA CCGAGGAGCT GCTGGTAAAG CTTAACAGAG
     3361 AAGATCTGTT GCGCAAACAG CGCACTTTCG ACAATGGAAG CATCCCCCAC CAGATTCACC
     3421 TGGGCGAACT GCACGCTATC CTCAGGCGGC AAGAGGATTT CTACCCCTTT TTGAAAGATA
     3481 ACAGGGAAAA GATTGAGAAA ATCCTCACAT TTCGGATACC CTACTATGTA GGCCCCCTCG
     3541 CCCGGGGAAA TTCCAGATTC GCGTGGATGA CTCGCAAATC AGAAGAGACt ATCACTCCCT
     3601 GGAACTTCGA GGAAGTCGTG GATAAGGGGG CCTCTGCCCA GTCCTTCATC GAAAGGATGA
     3661 CTAACTTTGA TAAAAATCTG CCTAACGAAA AGGTGCTTCC TAAACACTCT CTGCTGTACG
     3721 AGTACTTCAC AGTTTATAAC GAGCTCACCA AGGTCAAATA CGTCACAGAA GGGATGAGAA
     3781 AGCCAGCATT CCTGTCTGGA GAGCAGAAGA AAGCTATCGT GGACCTCCTC TTCAAGACGA
     3841 ACCGGAAAGT TACCGTGAAA CAGCTCAAAG AgGACTATTT CAAAAAGATT GAATGTTTCG
     3901 ACTCTGTTGA AATCAGCGGA GTGGAGGATC GCTTCAACGC ATCCCTGGGA ACGTATCACG
     3961 ATCTCCTGAA AATCATTAAA GACAAGGACT TCCTGGACAA TGAGGAGAAC GAGGACATTC
     4021 TTGAGGACAT TGTCCTCACC CTTACGTTGT TTGAAGATAG GGAGATGATT GAAGAACGCT
     4081 TGAAAACTTA CGCTCATCTC TTCGACGACA AAGTCATGAA ACAGCTCAAG AGGCGCCGAT
     4141 ATACAGGATG GGGGCGGCTG TCAAGAAAAC TGATCAATGG GATCCGAGAC AAGCAGAGTG
     4201 GAAAGACAAT CCTGGATTTT CTTAAGTCCG ATGGATTTGC CAACCGGAAC TTCATGCAGT
     4261 TGATCCATGA TGACTCTCTC ACCTTTAAGG AGGACATCCA GAAAGCACAA GTTTCTGGCC
     4321 AGGGGGACAG cCTTCACGAG CACATCGCTA ATCTTGCAGG TAGCCCAGCT ATCAAAAAGG
     4381 GAATACTGCA GACCGTTAAG GTCGTGGATG AACTCGTCAA AGTAATGGGA AGGCATAAGC
     4441 CCGAGAATAT CGTTATCGAG ATGGCCCGAG AGAACCAAAC TACCCAGAAG GGACAGAAGA
     4501 ACAGTAGGGA AAGGATGAAG AGGATTGAAG AGGGTATAAA AGAACTGGGG TCCCAAATCC
     4561 TTAAGGAACA CCCAGTTGAA AACACCCAGC TTCAGAATGA GAAGCTCTAC CTGTACTACC
     4621 TGCAGAACGG CAGGGACATG TACGTGGATC AGGAACTGGA CATCAATCGG CTCTCCGACT
     4681 ACGACGTGGA TCATATCGTG CCCCAGTCTT TTCTCAAAGA TGATTCTATT GATAATAAAG
     4741 TGTTGACAAG ATCCGATAAA AATAGAGGGA AGAGTGATAA CGTCCCCTCA GAAGAAGTTG
     4801 TCAAGAAAAT GAAAAATTAT TGGCGGCAGC TGCTGAACGC CAAACTGATC ACACAACGGA
     4861 AGTTCGATAA TCTGACTAAG GCTGAACGAG GTGGCCTGTC TGAGTTGGAT AAAGCCGGCT
     4921 TCATCAAAAG GCAGCTTGTT GAGACACGCC AGATCACCAA GCACGTGGCC CAAATTCTCG
     4981 ATTCACGCAT GAACACCAAG TACGATGAAA ATGACAAACT GATTCGAGAG GTGAAAGTTA
     5041 TTACTCTGAA GTCTAAGCTc GTCTCAGATT TCAGAAAGGA CTTTCAGTTT TATAAGGTGA
     5101 GAGAGATCAA CAATTACCAC CATGCGCATG ATGCCTACCT GAATGCAGTG GTAGGCACTG
     5161 CACTTATCAA AAAATATCCC AAGCTTGAAT CTGAATTTGT TTACGGAGAC TATAAAGTGT
     5221 ACGATGTTAG GAAAATGATC GCAAAGTCTG AGCAGGAAAT AGGCAAGGCC ACCGCTAAGT
     5281 ACTTCTTTTA CAGCAATATT ATGAATTTTT TCAAGACCGA GATTACACTG GCCAATGGAG
     5341 AGATTCGGAA GCGACCACTT ATCGAAACAA ACGGAGAAAC AGGAGAAATC GTGTGGGACA
     5401 AGGGTAGGGA TTTCGCGACA GTCCGGAAGG TCCTGTCCAT GCCGCAGGTG AACATCGTTA
     5461 AAAAGACCGA AGTACAGACC GGAGGCTTCT CCAAGGAAAG TATCCTCCCG AAAAGGAACA
     5521 GCGACAAGCT GATCGCACGC AAAAAAGATT GGGACCCCAA GAAATACGGC GGATTCGATT
     5581 CTCCTACAGT CGCTTACAGT GTACTGGTTG TGGCCAAAGT GGAGAAAGGG AAGTCTAAAA
     5641 AACTCAAAAG CGTCAAGGAA CTGCTGGGCA TCACAATCAT GGAGCGATCA AGCTTCGAAA
     5701 AAAACCCCAT CGACTTTCTC GAGGCGAAAG GATATAAAGA GGTCAAAAAA GACCTCATCA
     5761 TTAAGCTTCC CAAGTACTCT CTCTTTGAGC TTGAAAACGG CCGGAAACGA ATGCTCGCTA
     5821 GTGCGGGCGA GCTGCAGAAA GGTAACGAGC TGGCACTGCC CTCTAAATAC GTTAATTTCT
     5881 TGTATCTGGC CAGCCACTAT GAAAAGCTCA AAGGGTCcCC CGAAGATAAT GAGCAGAAGC
     5941 AGCTGTTCGT GGAACAACAC AAACACTACC TTGATGAGAT CATCGAGCAA ATAAGCGAAT
     6001 TCTCCAAAAG AGTGATCCTC GCCGACGCTA ACCTCGATAA GGTGCTTTCT GCTTACAATA
     6061 AGCACAGGGA TAAGCCCATC AGGGAGCAGG CAGAAAACAT TATCCACTTG TTTACTCTGA
     6121 CCAACTTGGG CGCGCCTGCA GCCTTCAAGT ACTTCGACAC CACCATAGAC AGAAAGCGGT
     6181 ACACCTCTAC AAAGGAGGTC CTGGACGCCA CACTGATTCA TCAGTCAATT ACGGGGCTCT
     6241 ATGAAACAAG AATCGACCTC TCTCAGCTCG GTGGAGACAG CAGGGCTGAC CCCAAGAAGA
     6301 AGAGGAAGGT GTGAGCTTCG TCCGACGGCG GCCCACGGGT CCCAGGCCTC GGAGATCCGT
     6361 CCCCCTTTTC CTTTGTCGAT ATCATGTAAT TAGTTATGTC ACGCTTACAT TCACGCCCTC
     6421 CCCCCACATC CGCTCTAACC GAAAAGGAAG GAGTTAGACA ACCTGAAGTC TAGGTCCCTA
     6481 TTTATTTTTT TATAGTTATG TTAGTATTAA GAACGTTATT TATATTTCAA ATTTTTCTTT
     6541 TTTTTCTGTA CAGACGCGTG TACGCATGTA ACATTATACT GAAAACCTTG CTTGAGAAGG
     6601 TTTTGGGACG CTCGAAGGCT TTAATTTGCA AGCTGGCGCT AATTCATGGA CATGGAGGCC
     6661 CAGAATACCC TCCTTGACAG TCTTGACGTG CGCAGCTCAG GGGCATGATG TGACTGTCGC
     6721 CCGTACATTT AGCCCATACA TCCCCATGTA TAATCATTTG CATCCATACA TTTTGATGGC
     6781 CGCACGGCGC GAAGCAAAAA TTACGGCTCC TCGCTGCAGA CGTGCGAGCA GGGAAACGCT
     6841 CCCCTCACAG ACGCGTTGAA TTGTCCCCAC GCCGCGCCCC TGTAGAGAAA TATAAAAGGT
     6901 TAGGATTTGC CACTGAGGTT CTTCTTTCAT ATACTTCCTT TTAAAATCTT GCTAGGATAC
     6961 AGTTCTCACA TCACATCCGA ACATAAACAA CCATGGGTAA GGAAAAGACT CACGTTTCGA
     7021 GGCCGCGATT AAATTCCAAC ATGGATGCTG ATTTATATGG GTATAAATGG GCTCGCGATA
     7081 ATGTCGGGCA ATCAGGTGCG ACAATCTATC GATTGTATGG GAAGCCCGAT GCGCCAGAGT
     7141 TGTTTCTGAA ACATGGCAAA GGTAGCGTTG CCAATGATGT TACAGATGAG ATGGTCAGAC
     7201 TAAACTGGCT GACGGAATTT ATGCCTCTTC CGACCATCAA GCATTTTATC CGTACTCCTG
     7261 ATGATGCATG GTTACTCACC ACTGCGATCC CCGGCAAAAC AGCATTCCAG GTATTAGAAG
     7321 AATATCCTGA TTCAGGTGAA AATATTGTTG ATGCGCTGGC AGTGTTCCTG CGCCGGTTGC
     7381 ATTCGATTCC TGTTTGTAAT TGTCCTTTTA ACAGCGATCG CGTATTTCGT CTCGCTCAGG
     7441 CGCAATCACG AATGAATAAC GGTTTGGTTG ATGCGAGTGA TTTTGATGAC GAGCGTAATG
     7501 GCTGGCCTGT TGAACAAGTC TGGAAAGAAA TGCATAAGCT TTTGCCATTC TCACCGGATT
     7561 CAGTCGTCAC TCATGGTGAT TTCTCACTTG ATAACCTTAT TTTTGACGAG GGGAAATTAA
     7621 TAGGTTGTAT TGATGTTGGA CGAGTCGGAA TCGCAGACCG ATACCAGGAT CTTGCCATCC
     7681 TATGGAACTG CCTCGGTGAG TTTTCTCCTT CATTACAGAA ACGGCTTTTT CAAAAATATG
     7741 GTATTGATAA TCCTGATATG AATAAATTGC AGTTTCATTT GATGCTCGAT GAGTTTTTCT
     7801 AATCAGTACT GACAATAAAA AGATTCTTGT TTTCAAGAAC TTGTCATTTG TATAGTTTTT
     7861 TTATATTGTA GTTGTTCTAT TTTAATCAAA TGTTAGCGTG ATTTATATTT TTTTTCGCCT
     7921 CGACATCATC TGCCCAGATG CGAAGTTAAG TGCGCAGAAA GTAATATCAT GCGTCAATCG
     7981 TATGTGAATG CTGGTCGCTA TACTGGCTTC CTTTTCATCA CGTGCTATAA AAATAATTAT
     8041 AATTTAAATT TTTTAATATA AATATATAAA TTAAAAATAG AAAGTAAAAA AAGAAATTAA
     8101 AGAAAAAATA GTTTTTGTTT TCCGAAGATG TAAAAGACTC TAGGGGGATC GCCAACAAAT
     8161 ACTACCTTTT ATCTTGCTCT TCCTGCTCTC AGGTATTAAT GCCGAATTGT TTCATCTTGT
     8221 CTGTGTAGAA CACCACACAC GAAAATCCTG TGATTTTACA TTTTACTTAT CGTTAATCGA
     8281 ATGTATATCT ATTTAATCTG CTTTTCTTGT CTAATAAATA TATATGTAAA GTACGCTTTT
     8341 TGTTGAAATT TTTTAAACCT TTGTTTATTT TTTTTTCTTC ATTCCGTAAC TCTTCTACCT
     8401 TCTTTATTTA CTTTCTAAAA TCCAAATACA AAACATAAAA ATAAATAAAC ACAGAGTAAA
     8461 TTCCCAAATT ATTCCATCAT TAAAAGATAC GAGGCGCGTG TAAGTTACAG GCA
//
