LOCUS       Mst1214_BB2_E_DE_pcoxA:pyrGtrunc_AMA1_2.8      7070 bp    DNA     circular UNA 
COMMENT     Note: the plasmid backbone was PCR amplified, only the linker sequence was checked by sequencing! (compare Seq folder)
FEATURES             Location/Qualifiers
     primer_bind     1..18
                     /label=BB2_seq_fw
     Vector          join(5018..7070,1..58)
                     /origin="PCRproductNewBB2 (2.125bp circular)Target
                     vector: from AgeI cut at 5^6 to SpeI cut at 2116^2117,
                     2.111bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     Vector          join(5002..7070,1..114)
                     /origin="BB2_L_DE_syn_BbsI_9 (2.215bp circular)Target
                     vector: from BbsI cut at 146^147 to BbsI cut at 114^115,
                     2.183bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     primer_bind     complement(41..57)
                     /label=BB2_BW
     Region          104..109
                     /label=BsaI
     FS              111..114
                     /label="FS D"
     primer          <115..>132
                     /label=FS12_empty(3)_fw
     FS              115..118
                     /label="FS 1"
     Region          115..>132
                     /note="Aspergillus niger chromosomal region withour known
                     function as spaver"
                     /label=Aniger_Chr1_54049_54070
     primer          complement(<119..>136)
                     /label=FS12_empty(3)_rev
     primer          <133..156
                     /label=FS2_pcoxA_BsaI_fw
     promoter        <133..1439
                     /label=pcoxA_199998
     primer          923..942
                     /label=2_coxA_fw
     primer          complement(923..942)
                     /label=2_coxA_rev
     mutation        933
                     /label=TamingT->A
     primer          complement(1193..1212)
                     /label=3_coxA_rev
     primer          1193..1212
                     /label=3_coxA_fw
     mutation        1202
                     /label=TamingA->T
     primer          complement(1422..>1443)
                     /label=FS2_pcoxA_rev
     FS              1440..1443
                     /label=FS2
     CDS             join(1441..1597,1598..>2119)
                     /source=AspGD
                     /ID=56726-mRNA-P
                     /Parent=56726-mRNA-T
                     /parent_feature_type=ORF
                     /frame="[complement(309053..309729):3,
                     complement(309799..309955): 0]"
                     /label=pyrGtrunc_56726-mRNA
     Mutated         complement(1445..1467)
                     /label="pyrG.1_binding site mutated"
     PAM             complement(1445..1447)
                     /label=pyrG.1_CRISPR
     Target          1487..1509
                     /label=pyrG.2_CRISPR
     PAM             1507..1509
                     /label=pyrG.2_CRISPR
     mutation        1536
                     /label=G>a
     mutation        1665
                     /label=C>t
     mutation        2073
                     /label=C>t
     primer          complement(2076..>2123)
                     /label=pyrG_trunc_FS3_BsaI_rev
     primer          <2120..2144
                     /label=FS3_AMA1/2_fw
     CDS             2124..5001
                     /label=AMA1/2
     primer          3090..3113
                     /label=1_AMA1/2_fw
     primer          complement(3090..3113)
                     /label=1_AMA1/2_rev
     Taming          3103
                     /label=A>t
     primer          complement(4979..>5005)
                     /label=FS4_AMA1/2_rev
     FS              5002..5005
                     /label="FS 4"
     FS              5006..5009
                     /label="FS E"
     Region          complement(5011..5016)
                     /label=BsaI
     primer_bind     5023..5038
                     /label=BB2_FW
     Region          complement(<5023..5039)
                     /label="f1 origin"
     primer_bind     complement(5114..5133)
                     /label=BB2_seq_bw
     CDS             5171..6028
                     /label=AmpicillinR
     misc_feature    5879..5895
                     /feature_type="Primer Region"
                     /label=BB2_BsaIMut_FW
     misc_feature    complement(5879..5895)
                     /feature_type="Primer Region"
                     /label=BB2_BsaIMut_BW
     primer          5887
                     /label="G changed to C"
     Region          6844..6901
                     /label="pUC origin"
     primer_bind     7002..7019
                     /label=pSTBlue_Seq_FW
ORIGIN
        1 CATTAATGCA GCTGGCACGA CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGCAACACT
       61 AGTcgcaaaa aaccccgccc ctgacagggc ggggtttttt cgcGGTCTCC AGCTGGAGAC
      121 GGGCACAAGG TTCATGAACC GAAGAGGATG GGAACCCCGA CCATGGCCTG CGGCCGCAGC
      181 TATCAGTCCT CTCAGCTGAC GCCCCTCACG AACCCTCGCA CTCTCCCGAA ATGCTCTTCG
      241 GTGTCCATGG CGCATCATCT CAGCAGCATC CCATGTCAAA CCATGGCTTT GGGCCCACGG
      301 ACTCTGTGGC CCTGCCGCAA CATCACCATC ACCATCGACT CCCGCCCCAT GCAGCGCTGC
      361 GCGCCCCAGG GCGTCATGGA GTGAATGTGG AATCTCCTCC TTTGCCCTCG GGCCCGCCGA
      421 GTGTGGTGGG CCAGCCTGGA ATGCCTGATC CAGCCCCCAG GCCTCGAGGA CCAAAACTGA
      481 AGTTTACTCC CGAAGAGGAC GCTCTACTGG TGGAGTTGAA GGAAAACAAG AACTTGACGT
      541 GGAAGCAAAT TGCAGACTTC TTCCCGGGCC GAACGAGCGG TACCTTGCAA GTCCGATACT
      601 GCACCAAGCT GAAGGCTAAG GATGTAGCTT GGAGTGACGA AATGGTTCGA TTTGCTCCTG
      661 ATGTATTTCT ACGCCTGTCT CACACATGCT AATGAAGGGA ATAGGTACAA AGGCTGCAGC
      721 GGGCAATGCA CGAGTACGAG AACGATCGGT GGCGCATCAT TGCAGGGAAG GTTGGAAATG
      781 GCTTCACCCC AGCTGCTTGC CGCGAGAAAG CCATGCAGCT CCATGAGTAA AAGCGTTGGG
      841 GAATTTTCAT ATTTATATCT ACTGTCGCCA GATTCGGCCC TGCTTGGACC CTCTGATCTC
      901 CTTACTCTCC ATATTGGTTC AAATGTCGGG TCACCGATAG GGCTGGTGGT GCAGGCTTGT
      961 TGTAGGCACG GGAGGATGAT CAGCATAACT CTGAGTCACT ATAGGGACGG GTTGATGTAA
     1021 GGTATTAAGT GATGTATGAT AATTCATTTT AGCCCGGGGG AACATATGGC GCCGGCATTT
     1081 GTTCGTTCGC AATGAACCGA CACTAGCGTC CGCTCTCGCA GTTTAGCACC GGCTGATCCC
     1141 GGGCTGAACG CGGCCATTGC TCGGCCGGGG CATGTGTTCC TTATCTACGG CAGACCGCAG
     1201 ATGACCACTG GAGCAGATTA TAGACCCTAA GCCCTAAGCC GGACACCCAA TCGAGTAGGT
     1261 CTGCGGACCA GGTCACTGCG GGCAGCCGGA GAAGCTCCGC AACCAATCAA TCCCCGGCGC
     1321 TGACTAAGGG CAGGCGACCA CGGGCCGAAG CGGCTTCAAA CTCACCTCAA CCTCCAAACT
     1381 CCCTCATCTC CAAACGTCCT TGCCTTGTCT GCCGTCATTG CAACCCACCC ACCAGGACAC
     1441 ATGAGCAGCA AGTCCCAGCT CACCTACACT GCCCGTGCCA GCAAGCATCC CAATGCTCTG
     1501 GCGAAGAGGC TGTTCGAGAT TGCCGAGGCC AAGAAaACCA ATGTGACTGT CTCGGCTGAC
     1561 GTTACCACCA CTAAGGAGCT ACTAGATCTT GCTGACCGTC TCGGTCCCTA CATTGCCGTG
     1621 ATCAAAACCC ACATCGATAT CCTCTCTGAT TTCAGCAACG AGACtATTGA GGGACTTAAG
     1681 GCTCTCGCGC AGAAGCACAA CTTTCTCATC TTCGAGGACC GCAAGTTCAT TGACATCGGC
     1741 AACACGGTCC AGAAGCAATA CCACGGCGGT ACCCTCCGTA TCTCGGAATG GGCCCACATC
     1801 ATCAACTGCA GCATTCTCCC TGGTGAGGGT ATCGTCGAGG CTCTCGCTCA GACGGCGTCT
     1861 GCACCGGACT TCGCCTACGG CCCCGAACGC GGTCTGTTGA TCTTGGCAGA GATGACCTCT
     1921 AAGGGCTCCT TGGCTACCGG CCAGTACACT ACTTCCTCGG TCGATTATGC CCGGAAATAC
     1981 AAGAACTTCG TTATGGGATT CGTGTCGACG CGCGCGTTGG GTGAGGTGCA GTCGGAAGTC
     2041 AGCTCTCCTT CGGATGAGGA GGACTTTGTG GTtTTCACGA CTGGTGTGAA CATTAACCTC
     2101 CAAGTCGCAA TTGACCTACG CTTATGCTGC AGACGCGTTA CGTATCGGAT CCAGAATTCG
     2161 TGATATCTGA ATTCGTCGAC AAGCTTATTT TTTGTATACT GTTTTGTGAT AGCACGAAGT
     2221 TTTTCCACGG TATCTTGTTA AAAATATATA TTTGTGGCGG GCTTACCTAC ATCAAATTAA
     2281 TAAGAGACTA ATTATAAACT AAACACACAA GCAAGCTACT TTAGGGTAAA AGTTTATAAA
     2341 TGCTTTTGAC GTATAAACGT TGCTTGTATT TATTATTACA ATTAAAGGTG GATAGAAAAC
     2401 CTAGAGACTA GTTAGAAACT AATCTCAGGT TTGCGTTAAA CTAAATCAGA GCCCGAGAGG
     2461 TTAACAGAAC CTAGAAGGGG ACTAGATATC CGGGTAGGGA AACAAAAAAA AAAAACAAGA
     2521 CAGCCACATA TTAGGGAGAC TAGTTAGAAG CTAGTTCCAG GACTAGGAAA ATAAAAGACA
     2581 ATGATACCAC AGTCTAGTTG ACAACTAGAT AGATTCTAGA TTGAGGCCAA AGTCTCTGAG
     2641 ATCCAGGTTA GTTGCAACTA ATACTAGTTA GTATCTAGTC TCCTATAACT CTGAAGCTAG
     2701 AATAACTTAC TACTATTATC CTCACCACTG TTCAGCTGCG CAAACGGAGT GATTGCAAGG
     2761 TGTTCAGAGA CTAGTTATTG ACTAGTCAGT GACTAGCAAT AACTAACAAG GTATTAACCT
     2821 ACCATGTCTG CCATCACCCT GCACTTCCTC GGGCTCAGCA GCCTTTTCCT CCTCATTTTC
     2881 ATGCTCATTT TCCTTGTTTA AGACTGTGAC TAGTCAAAGA CTAGTCCAGA ACCACAAAGG
     2941 AGAAATGTCT TACCACTTTC TTCATTGCTT GTCTCTTTTG CATTATCCAT GTCTGCAACT
     3001 AGTTAGAGTC TAGTTAGTGA CTAGTCCGAC GAGGACTTGC TTGTCTCCGG ATTGTTGGAG
     3061 GAACTCTCCA GGGCCTCAAG ATCCACAACA GAGCCTTCTA GAtGACTGGT CAATAACTAG
     3121 TTGGTCTTTG TCTGAGTCTG ACTGACTTAC GAGGTTGCAT ACTCGCTCCC TTTGCCTCGT
     3181 CAATCGATGA GAAAAAGCGC CAAAACTCGC AATATGGCTT TGAACCACAC GGTGCTGAGA
     3241 CTAGTTAGAA TCTAGTCCCA AACTAGCTTG GATAGCTTAC CTTTGCCCTT TGCGTTGCGA
     3301 CAGGTCTTGC AGGGTATGGT TCCTTTCTCA CCAGCTGATT TAGCTGCCTT GCTACCCTCA
     3361 CGGCGGATCT GCATAAAGAG TGGCTAGAGG TTATAAATTA GCACTGATCC TAGGTACGGG
     3421 GCTGAATGTA ACTTGCCTTT CCTTTCTCAT CGCGCGGCAA GACAGGCTTG CTCAAATTCC
     3481 TACCAGTCAC AGGGGTATGC ACGGCGTACG GACCACTTGA ACTAGTCACA GATTAGTTAG
     3541 CAACTAGTCT GCATTGAATG GCTGTACTTA CGGGCCCTCG CCATTGTCCT GATCATTTCC
     3601 AGCTTCACCC TCGTTGCTGC AAAGTAGTTA GTGACTAGTC AAGGACTAGT TGAAATGGGA
     3661 GAAGAAACTC ACGAATTCTC GACACCCTTA GTATTGTGGT CCTTGGACTT GGTGCTGCTA
     3721 TATATTAGCT AATACACTAG TTAGACTCAC AGAAACTTAC GCAGCTCGCT TGCGCTTCTT
     3781 GGTAGGAGTC GGGGTTGGGA GAACAGTGCC TTCAAACAAG CCTTCATACC ATGCTACTTG
     3841 ACTAGTCAGG GACTAGTCAC CAAGTAATCT AGATAGGACT TGCCTTTGGC CTCCATCAGT
     3901 TCCTTCATAG TGGGAGGTCC ATTGTGCAAT GTAAACTCCA TGCCGTGGGA GTTCTTGTCC
     3961 TTCAAGTGCT TGACCAATAT GTTTCTGTTG GCAGAGGGAA CCTGTCAACT AGTTAATAAC
     4021 TAGTCAGAAA CTAGTATAGC AGTAGACTCA CTGTACGCTT GAGGCATCCC TTCACTCGGC
     4081 AGTAGACTTC ATATGGATGG ATATCAGGCA CGCCATTGTC GTCCTGTGGA CTAGTCAGTA
     4141 ACTAGGCTTA AAGCTAGTCG GGTCGGCTTA CTATCTTGAA ATCCGGCAGC GTAAGCTCCC
     4201 CGTCCTTAAC TGCCTCGAGA TAGTGACAGT ACTCTGGGGA CTTTCGGAGA TCGTTATCGC
     4261 GAATGCTCGG CATACTAATC GTTGACTAGT CTTGGACTAG TCCCGAGCAA AAAGGATTGG
     4321 AGGAGGAGGA GGAAGGTGAG AGTGAGACAA AGAGCGAAAT AAGAGCTTCA AAGGCTATCT
     4381 CTAAGCAGTA TGAAGGTTAA GTATCTAGTT CTTGACTAGA TTTAAAAGAG ATTTCGACTA
     4441 GTTATGTACC TGGAGTTTGG ATATAGGAAT GTGTTGTGGT AACGAAATGT AAGGGGGAGG
     4501 AAAGAAAAAG TCGGTCAAGA GGTAACTCTA AGTCGGCCAT TCCTTTTTGG GAGGCGCTAA
     4561 CCATAAACGG CATGGTCGAC TTAGAGTTAG CTCAGGGAAT TTAGGGAGTT ATCTGCGACC
     4621 ACCGAGGAAC GGCGGAATGC CAAAGAATCC CGATGGAGCT CTAGCTGGCG GTTGACAACC
     4681 CCACCTTTTG GCGTTTCTGC GGCGTTGCAG GCGGGACTGG ATACTTCGTA GAACCAGAAA
     4741 GGCAAGGCAG AACGCGCTCA GCAAGAGTGT TGGAAGTGAT AGCATGATGT GCCTTGTTAA
     4801 CTAGGTCAAA ATCTGCAGTA TGCTTGATGT TATCCAAAGT GTGAGAGAGG AAGGTCCAAA
     4861 CATACACGAT TGGGAGAGGG CCTAGGTATA AGAGTTTTTG AGTAGAACGC ATGTGAGCCC
     4921 AGCCATCTCG AGGAGATTAA ACACGGGCCG GCATTTGATG GCTATGTTAG TACCCCAATG
     4981 GAAAGCCTGA GAGTCCAGTG GCGCTGGAGG GAGACCACCG GTGCCGCTAC AGGGCGCGTC
     5041 AGGTGGCACT TTTCGGGGAA ATGTGCGCGG AACCCCTATT TGTTTATTTT TCTAAATACA
     5101 TTCAAATATG TATCCGCTCA TGAGACAATA ACCCTGATAA ATGCTTCAAT AATATTGAAA
     5161 AAGGAAGAGT ATGAGTATTC AACATTTCCG TGTCGCCCTT ATTCCCTTTT TTGCGGCATT
     5221 TTGCCTTCCT GTTTTTGCTC ACCCAGAAAC GCTGGTGAAA GTAAAAGATG CTGAAGATCA
     5281 GTTGGGTGCA CGAGTGGGTT ACATCGAACT GGATCTCAAC AGCGGTAAGA TCCTTGAGAG
     5341 TTTTCGCCCC GAAGAACGTT TTCCAATGAT GAGCACTTTT AAAGTTCTGC TATGTGGCGC
     5401 GGTATTATCC CGTATTGACG CCGGGCAAGA GCAACTCGGT CGCCGCATAC ACTATTCTCA
     5461 GAATGACTTG GTTGAGTACT CACCAGTCAC AGAAAAGCAT CTTACGGATG GCATGACAGT
     5521 AAGAGAATTA TGCAGTGCTG CCATAACCAT GAGTGATAAC ACTGCGGCCA ACTTACTTCT
     5581 GACAACGATC GGAGGACCGA AGGAGCTAAC CGCTTTTTTG CACAACATGG GGGATCATGT
     5641 AACTCGCCTT GATCGTTGGG AACCGGAGCT GAATGAAGCC ATACCAAACG ACGAGCGTGA
     5701 CACCACGATG CCTGTAGCAA TGGCAACAAC GTTGCGCAAA CTATTAACTG GCGAACTACT
     5761 TACTCTAGCT TCCCGGCAAC AATTAATAGA CTGGATGGAG GCGGATAAAG TTGCAGGACC
     5821 ACTTCTGCGC TCGGCCCTTC CGGCTGGCTG GTTTATTGCT GATAAATCTG GAGCCGGTGA
     5881 GCGTGGCTCT CGCGGTATCA TTGCAGCACT GGGGCCAGAT GGTAAGCCCT CCCGTATCGT
     5941 AGTTATCTAC ACGACGGGGA GTCAGGCAAC TATGGATGAA CGAAATAGAC AGATCGCTGA
     6001 GATAGGTGCC TCACTGATTA AGCATTGGTA ACTGTCAGAC CAAGTTTACT CATATATACT
     6061 TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC TAGGTGAAGA TCCTTTTTGA
     6121 TAATCTCATG AACAATAAAA CTGTCTGCTT ACATAAACAG TAATACAAGG GGTGTAGATC
     6181 TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA
     6241 AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA
     6301 AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC
     6361 CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT
     6421 AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC
     6481 TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC
     6541 GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA
     6601 GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG
     6661 CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG
     6721 GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT
     6781 TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT
     6841 GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC
     6901 ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT
     6961 GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG
     7021 CGGAAGAGCG CCCAATACGC AAACCGCCTC TCCCCGCGCG TTGGCCGATT
//
