LOCUS       sequence-181729-        7066 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 7066)
  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 7066)
  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..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..43
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5159..5263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5264..5332,5333..6124)
                     /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(5482..5501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6350..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /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 tacgaggttg catactcgct ccctttgcct cgtcaatcga tgagaaaaag cgccaaaact
     3301 cgcaatatgg ctttgaacca cacggtgctg agactagtta gaatctagtc ccaaactagc
     3361 ttggatagct tacctttgcc ctttgcgttg cgacaggtct tgcagggtat ggttcctttc
     3421 tcaccagctg atttagctgc cttgctaccc tcacggcgga tctgcataaa gagtggctag
     3481 aggttataaa ttagcactga tcctaggtac ggggctgaat gtaacttgcc tttcctttct
     3541 catcgcgcgg caagacaggc ttgctcaaat tcctaccagt cacaggggta tgcacggcgt
     3601 acggaccact tgaactagtc acagattagt tagcaactag tctgcattga atggctgtac
     3661 ttacgggccc tcgccattgt cctgatcatt tccagcttca ccctcgttgc tgcaaagtag
     3721 ttagtgacta gtcaaggact agttgaaatg ggagaagaaa ctcacgaatt ctcgacaccc
     3781 ttagtattgt ggtccttgga cttggtgctg ctatatatta gctaatacac tagttagact
     3841 cacagaaact tacgcagctc gcttgcgctt cttggtagga gtcggggttg ggagaacagt
     3901 gccttcaaac aagccttcat accatgctac ttgactagtc agggactagt caccaagtaa
     3961 tctagatagg acttgccttt ggcctccatc agttccttca tagtgggagg tccattgtgc
     4021 aatgtaaact ccatgccgtg ggagttcttg tccttcaagt gcttgaccaa tatgtttctg
     4081 ttggcagagg gaacctgtca actagttaat aactagtcag aaactagtat agcagtagac
     4141 tcactgtacg cttgaggcat cccttcactc ggcagtagac ttcatatgga tggatatcag
     4201 gcacgccatt gtcgtcctgt ggactagtca gtaactaggc ttaaagctag tcgggtcggc
     4261 ttactatctt gaaatccggc agcgtaagct ccccgtcctt aactgcctcg agatagtgac
     4321 agtactctgg ggactttcgg agatcgttat cgcgaatgct cggcatacta atcgttgact
     4381 agtcttggac tagtcccgag caaaaaggat tggaggagga ggaggaaggt gagagtgaga
     4441 caaagagcga aataagagct tcaaaggcta tctctaagca gtatgaaggt taagtatcta
     4501 gttcttgact agatttaaaa gagatttcga ctagttatgt acctggagtt tggatatagg
     4561 aatgtgttgt ggtaacgaaa tgtaaggggg aggaaagaaa aagtcggtca agaggtaact
     4621 ctaagtcggc cattcctttt tgggaggcgc taaccataaa cggcatggtc gacttagagt
     4681 tagctcaggg aatttaggga gttatctgcg accaccgagg aacggcggaa tgccaaagaa
     4741 tcccgatgga gctctagctg gcggttgaca accccacctt ttggcgtttc tgcggcgttg
     4801 caggcgggac tggatacttc gtagaaccag aaaggcaagg cagaacgcgc tcagcaagag
     4861 tgttggaagt gatagcatga tgtgccttgt taactaggtc aaaatctgca gtatgcttga
     4921 tgttatccaa agtgtgagag aggaaggtcc aaacatacac gattgggaga gggcctaggt
     4981 ataagagttt ttgagtagaa cgcatgtgag cccagccatc tcgaggagat taaacacggg
     5041 ccggcatttg atggctatgt tagtacccca atggaaagcc tgagagtcca gtggcgctag
     5101 ctggagacca ccggtgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     5161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5941 ctggtttatt gctgataaat ctggagccgg tgagcgtggc tctcgcggta tcattgcagc
     6001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaacaata aaactgtctg
     6241 cttacataaa cagtaataca aggggtgtag atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6481 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgc
//
