LOCUS       sequence-176367-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 with split pyrG marker for homologous integration with linker CD
            (for 2 expression cassettes).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7070)
  AUTHORS   Sarkari P, Marx H, Blumhoff ML, Mattanovich D, Sauer M, Steiger MG
  TITLE     An efficient tool for metabolic pathway construction and gene 
            integration for Aspergillus niger.
  JOURNAL   Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004.
  PUBMED    28533066
REFERENCE   2  (bases 1 to 7070)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Bioresour 
            Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004."
COMMENT     Sequence Label: pMST1213_BB2_E_CD_pcoxA:pyrGtrunc_AMA1_2.8
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..43
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5163..5267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5268..5336,5337..6128)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5486..5505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6354..6942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6843..6862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
       61 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      121 gtttcccgac tggaaagcgg gcagtgagcg caacactagt cgcaaaaaac cccgcccctg
      181 acagggcggg gttttttcgc ggtctccaat tggagacggg cacaaggttc atgaaccgaa
      241 gaggatggga accccgacca tggcctgcgg ccgcagctat cagtcctctc agctgacgcc
      301 cctcacgaac cctcgcactc tcccgaaatg ctcttcggtg tccatggcgc atcatctcag
      361 cagcatccca tgtcaaacca tggctttggg cccacggact ctgtggccct gccgcaacat
      421 caccatcacc atcgactccc gccccatgca gcgctgcgcg ccccagggcg tcatggagtg
      481 aatgtggaat ctcctccttt gccctcgggc ccgccgagtg tggtgggcca gcctggaatg
      541 cctgatccag cccccaggcc tcgaggacca aaactgaagt ttactcccga agaggacgct
      601 ctactggtgg agttgaagga aaacaagaac ttgacgtgga agcaaattgc agacttcttc
      661 ccgggccgaa cgagcggtac cttgcaagtc cgatactgca ccaagctgaa ggctaaggat
      721 gtagcttgga gtgacgaaat ggttcgattt gctcctgatg tatttctacg cctgtctcac
      781 acatgctaat gaagggaata ggtacaaagg ctgcagcggg caatgcacga gtacgagaac
      841 gatcggtggc gcatcattgc agggaaggtt ggaaatggct tcaccccagc tgcttgccgc
      901 gagaaagcca tgcagctcca tgagtaaaag cgttggggaa ttttcatatt tatatctact
      961 gtcgccagat tcggccctgc ttggaccctc tgatctcctt actctccata ttggttcaaa
     1021 tgtcgggtca ccgatagggc tggtggtgca ggcttgttgt aggcacggga ggatgatcag
     1081 cataactctg agtcactata gggacgggtt gatgtaaggt attaagtgat gtatgataat
     1141 tcattttagc ccgggggaac atatggcgcc ggcatttgtt cgttcgcaat gaaccgacac
     1201 tagcgtccgc tctcgcagtt tagcaccggc tgatcccggg ctgaacgcgg ccattgctcg
     1261 gccggggcat gtgttcctta tctacggcag accgcagatg accactggag cagattatag
     1321 accctaagcc ctaagccgga cacccaatcg agtaggtctg cggaccaggt cactgcgggc
     1381 agccggagaa gctccgcaac caatcaatcc ccggcgctga ctaagggcag gcgaccacgg
     1441 gccgaagcgg cttcaaactc acctcaacct ccaaactccc tcatctccaa acgtccttgc
     1501 cttgtctgcc gtcattgcaa cccacccacc aggacacatg agcagcaagt cccagctcac
     1561 ctacactgcc cgtgccagca agcatcccaa tgctctggcg aagaggctgt tcgagattgc
     1621 cgaggccaag aaaaccaatg tgactgtctc ggctgacgtt accaccacta aggagctact
     1681 agatcttgct gaccgtctcg gtccctacat tgccgtgatc aaaacccaca tcgatatcct
     1741 ctctgatttc agcaacgaga ctattgaggg acttaaggct ctcgcgcaga agcacaactt
     1801 tctcatcttc gaggaccgca agttcattga catcggcaac acggtccaga agcaatacca
     1861 cggcggtacc ctccgtatct cggaatgggc ccacatcatc aactgcagca ttctccctgg
     1921 tgagggtatc gtcgaggctc tcgctcagac ggcgtctgca ccggacttcg cctacggccc
     1981 cgaacgcggt ctgttgatct tggcagagat gacctctaag ggctccttgg ctaccggcca
     2041 gtacactact tcctcggtcg attatgcccg gaaatacaag aacttcgtta tgggattcgt
     2101 gtcgacgcgc gcgttgggtg aggtgcagtc ggaagtcagc tctccttcgg atgaggagga
     2161 ctttgtggtt ttcacgactg gtgtgaacat taacctccaa gtcgcaattg acctacgctt
     2221 atgctgcaga cgcgttacgt atcggatcca gaattcgtga tatctgaatt cgtcgacaag
     2281 cttatttttt gtatactgtt ttgtgatagc acgaagtttt tccacggtat cttgttaaaa
     2341 atatatattt gtggcgggct tacctacatc aaattaataa gagactaatt ataaactaaa
     2401 cacacaagca agctacttta gggtaaaagt ttataaatgc ttttgacgta taaacgttgc
     2461 ttgtatttat tattacaatt aaaggtggat agaaaaccta gagactagtt agaaactaat
     2521 ctcaggtttg cgttaaacta aatcagagcc cgagaggtta acagaaccta gaaggggact
     2581 agatatccgg gtagggaaac aaaaaaaaaa aacaagacag ccacatatta gggagactag
     2641 ttagaagcta gttccaggac taggaaaata aaagacaatg ataccacagt ctagttgaca
     2701 actagataga ttctagattg aggccaaagt ctctgagatc caggttagtt gcaactaata
     2761 ctagttagta tctagtctcc tataactctg aagctagaat aacttactac tattatcctc
     2821 accactgttc agctgcgcaa acggagtgat tgcaaggtgt tcagagacta gttattgact
     2881 agtcagtgac tagcaataac taacaaggta ttaacctacc atgtctgcca tcaccctgca
     2941 cttcctcggg ctcagcagcc ttttcctcct cattttcatg ctcattttcc ttgtttaaga
     3001 ctgtgactag tcaaagacta gtccagaacc acaaaggaga aatgtcttac cactttcttc
     3061 attgcttgtc tcttttgcat tatccatgtc tgcaactagt tagagtctag ttagtgacta
     3121 gtccgacgag gacttgcttg tctccggatt gttggaggaa ctctccaggg cctcaagatc
     3181 cacaacagag ccttctagat gactggtcaa taactagttg gtctttgtct gagtctgact
     3241 gacttacgag gttgcatact cgctcccttt gcctcgtcaa tcgatgagaa aaagcgccaa
     3301 aactcgcaat atggctttga accacacggt gctgagacta gttagaatct agtcccaaac
     3361 tagcttggat agcttacctt tgccctttgc gttgcgacag gtcttgcagg gtatggttcc
     3421 tttctcacca gctgatttag ctgccttgct accctcacgg cggatctgca taaagagtgg
     3481 ctagaggtta taaattagca ctgatcctag gtacggggct gaatgtaact tgcctttcct
     3541 ttctcatcgc gcggcaagac aggcttgctc aaattcctac cagtcacagg ggtatgcacg
     3601 gcgtacggac cacttgaact agtcacagat tagttagcaa ctagtctgca ttgaatggct
     3661 gtacttacgg gccctcgcca ttgtcctgat catttccagc ttcaccctcg ttgctgcaaa
     3721 gtagttagtg actagtcaag gactagttga aatgggagaa gaaactcacg aattctcgac
     3781 acccttagta ttgtggtcct tggacttggt gctgctatat attagctaat acactagtta
     3841 gactcacaga aacttacgca gctcgcttgc gcttcttggt aggagtcggg gttgggagaa
     3901 cagtgccttc aaacaagcct tcataccatg ctacttgact agtcagggac tagtcaccaa
     3961 gtaatctaga taggacttgc ctttggcctc catcagttcc ttcatagtgg gaggtccatt
     4021 gtgcaatgta aactccatgc cgtgggagtt cttgtccttc aagtgcttga ccaatatgtt
     4081 tctgttggca gagggaacct gtcaactagt taataactag tcagaaacta gtatagcagt
     4141 agactcactg tacgcttgag gcatcccttc actcggcagt agacttcata tggatggata
     4201 tcaggcacgc cattgtcgtc ctgtggacta gtcagtaact aggcttaaag ctagtcgggt
     4261 cggcttacta tcttgaaatc cggcagcgta agctccccgt ccttaactgc ctcgagatag
     4321 tgacagtact ctggggactt tcggagatcg ttatcgcgaa tgctcggcat actaatcgtt
     4381 gactagtctt ggactagtcc cgagcaaaaa ggattggagg aggaggagga aggtgagagt
     4441 gagacaaaga gcgaaataag agcttcaaag gctatctcta agcagtatga aggttaagta
     4501 tctagttctt gactagattt aaaagagatt tcgactagtt atgtacctgg agtttggata
     4561 taggaatgtg ttgtggtaac gaaatgtaag ggggaggaaa gaaaaagtcg gtcaagaggt
     4621 aactctaagt cggccattcc tttttgggag gcgctaacca taaacggcat ggtcgactta
     4681 gagttagctc agggaattta gggagttatc tgcgaccacc gaggaacggc ggaatgccaa
     4741 agaatcccga tggagctcta gctggcggtt gacaacccca ccttttggcg tttctgcggc
     4801 gttgcaggcg ggactggata cttcgtagaa ccagaaaggc aaggcagaac gcgctcagca
     4861 agagtgttgg aagtgatagc atgatgtgcc ttgttaacta ggtcaaaatc tgcagtatgc
     4921 ttgatgttat ccaaagtgtg agagaggaag gtccaaacat acacgattgg gagagggcct
     4981 aggtataaga gtttttgagt agaacgcatg tgagcccagc catctcgagg agattaaaca
     5041 cgggccggca tttgatggct atgttagtac cccaatggaa agcctgagag tccagtggcg
     5101 ctagctggag accaccggtg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     5161 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5221 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5281 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5341 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5401 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5461 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5521 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5581 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5641 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5701 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5761 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5821 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5881 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5941 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg
     6001 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6061 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6121 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6181 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgaac aataaaactg
     6241 tctgcttaca taaacagtaa tacaaggggt gtagatctca tgaccaaaat cccttaacgt
     6301 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6361 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6421 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6481 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6541 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6601 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6661 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6721 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6781 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6841 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6901 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6961 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7021 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
//
