LOCUS       BB3_L_hygro_2µ_MST1312_ptpi1_TMB_tcyc_URA3_PT      7519 bp    DNA     circular UNA 
FEATURES             Location/Qualifiers
     primer          complement(join(7502..7519,1..>4))
                     /label="FS4_2micronBW"
     Vector          1..678
                     /origin="pSTBLUE-1 wo_ORI_circular_pUC_ORI_aus_pGAPZB
                     (2.745bp circular)Target vector: from BbsI cut at
                     2054^2055 to BbsI cut at 2732^2733, 678bpOrientation:
                     originalPlus strand 5':   no changePlus strand 3':   no
                     changeMinus strand 5':  no changeMinus strand 3':  no
                     change"
     Vector          join(4460..7519,1..722)
                     /origin="BB3_L_hygro_2µ_AD (3,812bp circular)Target
                     vector: from BsaI cut at 752^753 to BsaI cut at 722^723,
                     3,782bpOrientation: 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"
     terminator      683..722
                     /label="BBa_B1007"
     FS              723..726
                     /label="FS A"
     Vector          join(3278..>3281,<723..726)
                     /origin="BB2_L_AB_syn_BbsI_6 (2,215bp circular)Target
                     vector: from BpiI cut at 273^274 to BpiI cut at 241^242,
                     2,183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     misc_feature    727..1379
                     /feature_type="Insertion"
                     /origin="BB1_P_12_sce_pTPI1_47 (3,589bp
                     circular)Fragment: from BpiI cut at 2149^2150 to BpiI cut
                     at 2802^2803, 653bpOrientation: originalPlus strand 5':
                     no changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
                     /label="pTPI1"
     Region          <727..>1379
                     /label="FS1_TPIpt_FS2"
     misc_feature    <1380..1383
                     /feature_type="Putative promoter 1000bp"
                     /source="AspGD"
                     /ID="57285-mRNA"
     primer          <1380..1404
                     /label="F2(BsaI)_TMB_(new)fw"
     FS              1380..1383
                     /label="FS2"
     CDS             join(1381..1641,1642..1882,1883..1951,1952..2398,2399..2955)
                     /source="AspGD"
                     /ID="57285-mRNA-P"
                     /Parent="57285-mRNA-T"
                     /parent_feature_type="ORF"
                     /frame="[721357..721617: 0, 721675..721915: 1,
                     721965..722014: 0, 722101..722530: 0, 722590..723146: 2]"
                     /label="57285-mRNA"
     gene            join(<1381..>1641,<1642..>1882,<1883..>1951,<1952..>2398,<2399..2955)
                     /source="AspGD"
                     /ID="57285-mRNA"
     mutation        1386
                     /label="A>c"
     mutation        1398
                     /label="A>c"
     cRNA            complement(1519..1541)
                     /label="TMB_1"
     primer          1532..1556
                     /label="2_TMB_fw"
     primer          complement(1532..1556)
                     /label="2_TMB_rev"
     mutation        1542
                     /label="G>a"
     cRNA            complement(1574..1596)
                     /label="TMB_2"
     primer          2087..2115
                     /label="3_TMB_fw"
     primer          complement(2087..2115)
                     /label="3_TMB_rev"
     mutation        2097
                     /label="A>g"
     primer          2179..2195
                     /label="4_TMB_fw"
     primer          complement(2179..2195)
                     /label="4_TMB_rev"
     primer          complement(2933..>2959)
                     /label="FS3_(bsaI)_TMB_rev"
     terminator      <2956..>3277
                     /label="tCYC1"
     FS              3278..3281
                     /label="FS 4"
     misc_feature    join(<4416..>4419,<3282..>3285)
                     /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(4416..>4419,<3282..3285)
                     /origin="BB2_BsaI_B_1_4_C_FW (2.215bp circular)Target
                     vector: from BpiI cut at 2199^2200 to BpiI cut at
                     2167^2168, 2.183bpOrientation: originalPlus strand 5':
                     no changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer          <3286..3304
                     /label="URA3_marker_ampl_fw1_BpiI_FS1"
     promoter        3290..3522
                     /label="URA3 promoter acc pRS"
     CDS             3523..4326
                     /label="Ura3p"
     gene            3523..4326
                     /score="0.99"
                     /source="AUGUSTUS"
                     /ID="g2024"
                     /Product="Ura3p"
                     /Note="corresponds to s288c YEL021W"
     gene            complement(3558..4034)
                     /source="AUGUSTUS"
                     /ID="g2025"
                     /Product="Ura3p"
                     /Note="corresponds to s288c YEL021W"
     primer          3774..3808
                     /label="URA3_marker_mut_fw2"
     primer          complement(3774..3808)
                     /label="URA3_marker_mut_rev2"
     Site            3792
                     /site_type="mutagenized site"
                     /label="A->G"
     primer          4180..4209
                     /label="URA3_marker_mut_fw3"
     primer          complement(4180..4209)
                     /label="URA3_marker_mut_rev3"
     Site            4194
                     /site_type="mutagenized site"
                     /label="C->G"
     terminator      4327..4415
                     /label="Terminator Region"
     primer          complement(4403..>4419)
                     /label="URA3_marker_ampl_rev1_BpiI_FS4"
     CDS             4468..4811
                     /label="TEFPromoter"
     misc_feature    4641
                     /label="C to G"
     CDS             4812..5840
                     /label="hphR"
     misc_feature    5123
                     /label="G to C"
     CDS             5849..6043
                     /label="TEFterminator"
     primer          <6044..6070
                     /label="FS3_2micronFW"
     oriT            <6044..7519
                     /label="2 micron (µ) origin"
     primer          6048..6070
                     /label="2micronFW"
     primer          6505..6525
                     /label="2micron Inner1FW"
     primer          complement(6505..6525)
                     /label="2micron Inner1BW"
     modified_base   6517
                     /label="C changed to G"
     primer          complement(7502..7519)
                     /label="2micronBW"
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 TTCTCAATGC 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 GAATCCATCA ATAGATACGT CCTGAGGACC GTGCTACCCA AATGGACTGA
      781 TTGTGAGGGA GAgCTAACTA CATAGTGTTT AAAGATTACG GATATTTAAC TTACTTAGAA
      841 TAATGCCATT TTTTTGAGTT ATAATAATCC TACGTTAGTG TGAGCGGGAT TTAAACTGTG
      901 AGGACCTCAA TACATTCAGA CACTTCTGAC GGTATCACCC TACTTATTCC CTTCGAGATT
      961 ATATCTAGGA ACCCATCAGG TTGGTGGAAG ATTACCCGTT CTAAGACTTT TCAGCTTCCT
     1021 CTATTGATGT TACACTCGGA CACCCCTTTT CTGGCATCCA GTTTTTAATC TTCAGTGGCA
     1081 TGTGAGATTC TCCGAAATTA ATTAAAGCAA TCACACAATT CTCTCGGATA CCACCTCGGT
     1141 TGAAACTGAC AGGTGGTTTG TTACGCATGC TAATGCAAAG GAGCCTATAT ACCTTTGGCT
     1201 CGGCTGCTGT AACAGGGAAT ATAAAGGGCA GCATAATTTA GGAGTTTAGT GAACTTGCAA
     1261 CATTTACTAT TTTCCCTTCT TACGTAAATA TTTTTCTTTT TAATTCTAAA TCAATCTTTT
     1321 TCAATTTTTT GTTTGTATTC TTTTCTTGCT TAAATCTATA ACTACAAAAA ACACATACAC
     1381 ATGTCcTCAA CCACGTCcTC ATCAAGATCA GACCTTGAAA AGGTCCCCGT ACCACAGGTC
     1441 ACCCCTAGAG ACAGTGACTC CGATAAGGGA TCCCTCTCTC CGGAGCCTTC GACCCTCGAG
     1501 GCTCAGTCAT CCGAGAAGCC ACCGCATCAT ATCTTCACAC GaTCTCGCAA GCTGCAAATG
     1561 GTTTGCATCG TCTCCCTCGC TGCCATATTT TCTCCGCTTT CGTCGAACAT TTACTTCCCT
     1621 GCCCTGGATG ATGTCTCGAA ATCCCTCAAC ATCAGCATGT CGCTCGCAAC ACTCACCATC
     1681 ACGGTGTACA TGATCGTCCA AGGCCTCGCT CCCAGCTTCT GGGGTTCCAT GTCAGACGCC
     1741 ACAGGTAGAC GGCCTGTCTT TATTGGAACA TTCATTGTTT ACCTCGTAGC CAATATTGCT
     1801 CTGGCCGAAT CCAAGAACTA TGGTGAGCTC ATGGCCTTCC GAGCCTTGCA GGCTGCTGGT
     1861 AGCGCAGCCA CCATCTCAAT CGGAGCTGGA GTGATTGGTG ATATCACAAA CTCGGAAGAA
     1921 AGAGGTAGCT TGGTGGGTAT CTTCGGTGGA GTTCGCATGC TTGGACAGGG AATCGGGCCG
     1981 GTTTTCGGCG GCATTTTCAC CCAGTATCTC GGATATCGAT CTATCTTTTG GTTCCTCACG
     2041 ATTGCTGGAG GCGTGAGTCT CCTGTCCATT CTGGTGCTTC TTCCGGAGAC ATTGAGgCCA
     2101 ATTGCTGGAA ATGGAACTGT GAAGCTCAAT GGCATTCATA AGCCCTTCAT CTACACGATC
     2161 ACCGGCCAGA CGGGGGTTGT CGAGGGAGCG CAACCGGAAG CGAAAAAGAC CAAAACCAGC
     2221 TGGAAGTCTG TTTTTGCTCC TTTGACATTC CTCGTCGAAA AGGACGTTTT CATCACCCTG
     2281 TTCTTTGGAA GTATCGTGTA CACAGTGTGG AGCATGGTGA CATCCAGTAC CACCGACCTC
     2341 TTCAGCGAAG TGTACGGCCT GTCATCCCTG GACATTGGAC TCACTTTCCT AGGCAATGGC
     2401 TTTGGATGTA TGTCTGGCTC TTATCTGGTC GGCTACCTTA TGGATTACAA CCACCGTCTT
     2461 ACCGAACGCG AATATTGCGA GAAACACGGT TATCCGGCAG GCACACGTGT CAATCTGAAA
     2521 TCACACCCCG ACTTCCCCAT TGAGGTCGCC CGGATGCGCA ATACCTGGTG GGTGATTGCG
     2581 ATCTTCATCG TGACAGTTGC TTTGTACGGC GTGTCTTTGC GGACACATCT GGCGGTGCCT
     2641 ATCATTCTGC AGTACTTCAT TGCGTTCTGC TCAACAGGAC TCTTCACCAT CAACAGCGCC
     2701 CTGGTCATCG ATCTTTACCC AGGTGCTAGC GCCAGTGCGA CAGCAGTGAA CAATCTGATG
     2761 CGGTGCCTGC TTGGAGCTGG CGGTGTGGCT ATCGTGCAAC CTATCCTGGA CGCCTTGAAG
     2821 CCGGATTATA CTTTCCTCTT GCTTGCCGGC ATCACCCTTG TGATGACTCC GTTGCTGTAC
     2881 GTCGAAGATC GATGGGGTCC TGGCTGGCGA CATGCCCGCG AAAGGAGACT CAAGGCCAAA
     2941 GCCAACGGCA ACTAGGCTTC GTCCGACGGC GGCCCACGGG TCCCAGGCCT CGGAGATCCG
     3001 TCCCCCTTTT CCTTTGTCGA TATCATGTAA TTAGTTATGT CACGCTTACA TTCACGCCCT
     3061 CCCCCCACAT CCGCTCTAAC CGAAAAGGAA GGAGTTAGAC AACCTGAAGT CTAGGTCCCT
     3121 ATTTATTTTT TTATAGTTAT GTTAGTATTA AGAACGTTAT TTATATTTCA AATTTTTCTT
     3181 TTTTTTCTGT ACAGACGCGT GTACGCATGT AACATTATAC TGAAAACCTT GCTTGAGAAG
     3241 GTTTTGGGAC GCTCGAAGGC TTTAATTTGC AAGCTGGCGC TCCGGGGAGC AGGGTCCATA
     3301 AAGCTTTTCA ATTCATCTTT TTTTTTTTAT TCTTTTTTTT GATTCCGGTT TCCTTGAAAT
     3361 TTTTTTGATT CGGTAATCTC CGAGCAGAAG GAAGAACGAA GGAAGGAGCA CAGACTTAGA
     3421 TTGGTATATA TACGCATATG TAGTGTTGAA GAAACATGAA ATTGCCCAGT ATTCTTAACC
     3481 CAACTGCACA GAACAAAAAC CTGCAGGAAA CGAAGATAAA TCATGTCGAA AGCTACATAT
     3541 AAGGAACGTG CTGCTACTCA TCCTAGTCCT GTTGCTGCCA AGCTATTTAA TATCATGCAC
     3601 GAAAAGCAAA CAAACTTGTG TGCTTCATTG GATGTTCGTA CCACCAAGGA ATTACTGGAG
     3661 TTAGTTGAAG CATTAGGTCC CAAAATTTGT TTACTAAAAA CACATGTGGA TATCTTGACT
     3721 GATTTTTCCA TGGAGGGCAC AGTTAAGCCG CTAAAGGCAT TATCCGCCAA GTACAATTTT
     3781 TTACTCTTCG AgGACAGAAA ATTTGCTGAC ATTGGTAATA CAGTCAAATT GCAGTACTCT
     3841 GCGGGTGTAT ACAGAATAGC AGAATGGGCA GACATTACGA ATGCACACGG TGTGGTGGGC
     3901 CCAGGTATTG TTAGCGGTTT GAAGCAGGCG GCGGAAGAAG TAACAAAGGA ACCTAGAGGC
     3961 CTTTTGATGT TAGCAGAATT GTCATGCAAG GGCTCCCTAG CTACTGGAGA ATATACTAAG
     4021 GGTACTGTTG ACATTGCGAA GAGCGACAAA GATTTTGTTA TCGGCTTTAT TGCTCAAAGA
     4081 GACATGGGTG GAAGAGATGA AGGTTACGAT TGGTTGATTA TGACACCCGG TGTGGGTTTA
     4141 GATGACAAGG GAGACGCATT GGGTCAACAG TATAGAACCG TGGATGATGT GGTgTCTACA
     4201 GGATCTGACA TTATTATTGT TGGAAGAGGA CTATTTGCAA AGGGAAGGGA TGCTAAGGTA
     4261 GAGGGTGAAC GTTACAGAAA AGCAGGCTGG GAAGCATATT TGAGAAGATG CGGCCAGCAA
     4321 AACTAAAAAA CTGTATTATA AGTAAATGCA TGTATACTAA ACTCACAAAT TAGAGCTTCA
     4381 ATTTAATTAT ATCAGTTATT ACCCGGGAAT CTCGGCGCTA ATTGGAGGCG TCTTCGGATC
     4441 CCTCGAGGAA GACGCCGCTA GCTCATGGAC ATGGAGGCCC AGAATACCCT CCTTGACAGT
     4501 CTTGACGTGC GCAGCTCAGG GGCATGATGT GACTGTCGCC CGTACATTTA GCCCATACAT
     4561 CCCCATGTAT AATCATTTGC ATCCATACAT TTTGATGGCC GCACGGCGCG AAGCAAAAAT
     4621 TACGGCTCCT CGCTGCAGAC GTGCGAGCAG GGAAACGCTC CCCTCACAGA CGCGTTGAAT
     4681 TGTCCCCACG CCGCGCCCCT GTAGAGAAAT ATAAAAGGTT AGGATTTGCC ACTGAGGTTC
     4741 TTCTTTCATA TACTTCCTTT TAAAATCTTG CTAGGATACA GTTCTCACAT CACATCCGAA
     4801 CATAAACAAC CATGGGTAAA AAGCCTGAAC TCACCGCGAC GTCTGTCGAG AAGTTTCTGA
     4861 TCGAAAAGTT CGACAGCGTC TCCGACCTGA TGCAGCTCTC GGAGGGCGAA GAATCTCGTG
     4921 CTTTCAGCTT CGATGTAGGA GGGCGTGGAT ATGTCCTGCG GGTAAATAGC TGCGCCGATG
     4981 GTTTCTACAA AGATCGTTAT GTTTATCGGC ACTTTGCATC GGCCGCGCTC CCGATTCCGG
     5041 AAGTGCTTGA CATTGGGGAA TTCAGCGAGA GCCTGACCTA TTGCATCTCC CGCCGTGCAC
     5101 AGGGTGTCAC GTTGCAAGAC CTCCCTGAAA CCGAACTGCC CGCTGTTCTG CAGCCGGTCG
     5161 CGGAGGCCAT GGATGCGATC GCTGCGGCCG ATCTTAGCCA GACGAGCGGG TTCGGCCCAT
     5221 TCGGACCGCA AGGAATCGGT CAATACACTA CATGGCGTGA TTTCATATGC GCGATTGCTG
     5281 ATCCCCATGT GTATCACTGG CAAACTGTGA TGGACGACAC CGTCAGTGCG TCCGTCGCGC
     5341 AGGCTCTCGA TGAGCTGATG CTTTGGGCCG AGGACTGCCC CGAAGTCCGG CACCTCGTGC
     5401 ACGCGGATTT CGGCTCCAAC AATGTCCTGA CGGACAATGG CCGCATAACA GCGGTCATTG
     5461 ACTGGAGCGA GGCGATGTTC GGGGATTCCC AATACGAGGT CGCCAACATC TTCTTCTGGA
     5521 GGCCGTGGTT GGCTTGTATG GAGCAGCAGA CGCGCTACTT CGAGCGGAGG CATCCGGAGC
     5581 TTGCAGGATC GCCGCGGCTC CGGGCGTATA TGCTCCGCAT TGGTCTTGAC CAACTCTATC
     5641 AGAGCTTGGT TGACGGCAAT TTCGATGATG CAGCTTGGGC GCAGGGTCGA TGCGACGCAA
     5701 TCGTCCGATC CGGAGCCGGG ACTGTCGGGC GTACACAAAT CGCCCGCAGA AGCGCGGCCG
     5761 TCTGGACCGA TGGCTGTGTA GAAGTACTCG CCGATAGTGG AAACCGACGC CCCAGCACTC
     5821 GTCCGAGGGC AAAGGAATAA TCAGTACTGA CAATAAAAAG ATTCTTGTTT TCAAGAACTT
     5881 GTCATTTGTA TAGTTTTTTT ATATTGTAGT TGTTCTATTT TAATCAAATG TTAGCGTGAT
     5941 TTATATTTTT TTTCGCCTCG ACATCATCTG CCCAGATGCG AAGTTAAGTG CGCAGAAAGT
     6001 AATATCATGC GTCAATCGTA TGTGAATGCT GGTCGCTATA CTGGCTTtta tcgatgataa
     6061 gctgtcaaag atgagaatta attccacgga ctatagacta tactagatac tccgtctact
     6121 gtacgataca cttccgctca ggtccttgtc ctttaacgag gccttaccac tcttttgtta
     6181 ctctattgat ccagctcagc aaaggcagtg tgatctaaga ttctatcttc gcgatgtagt
     6241 aaaactagct agaccgagaa agagactaga aatgcaaaag gcacttctac aatggctgcc
     6301 atcattatta tccgatgtga cgctgcagct tctcaatgat attcgaatac gctttgagga
     6361 gatacagcct aatatccgac aaactgtttt acagatttac gatcgtactt gttacccatc
     6421 attgaatttt gaacatccga acctgggagt tttccctgaa acagatagta tatttgaacc
     6481 tgtataataa tatatagtct agcgctttac ggaagagaat gtatgtattt cggttcctgg
     6541 agaaactatt gcatctattg cataggtaat cttgcacgtc gcatccccgg ttcattttct
     6601 gcgtttccat cttgcacttc aatagcatat ctttgttaac gaagcatctg tgcttcattt
     6661 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     6721 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     6781 atttttgtaa aacaaaaatg caacgcgacg agagcgctaa tttttcaaac aaagaatctg
     6841 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     6901 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     6961 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     7021 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     7081 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     7141 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     7201 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     7261 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     7321 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     7381 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     7441 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     7501 ggaagcggta ttcgcaatg
//
