LOCUS       sequence-176369-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 with split pyrG marker for homologous integration with linker AB
            (negative control).
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: pMST1211_BB2_E_AB_pcoxA:pyrGtrunc_AMA1_2.8
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     326..345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     579..596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5716..5820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5821..5889,5890..6681)
                     /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(6039..6058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6907..7070,1..425)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
       61 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      121 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      181 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      241 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      301 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      361 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      421 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      481 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      541 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      601 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      661 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacact agtcgcaaaa
      721 aaccccgccc ctgacagggc ggggtttttt cgcggtctcc gatcggagac gggcacaagg
      781 ttcatgaacc gaagaggatg ggaaccccga ccatggcctg cggccgcagc tatcagtcct
      841 ctcagctgac gcccctcacg aaccctcgca ctctcccgaa atgctcttcg gtgtccatgg
      901 cgcatcatct cagcagcatc ccatgtcaaa ccatggcttt gggcccacgg actctgtggc
      961 cctgccgcaa catcaccatc accatcgact cccgccccat gcagcgctgc gcgccccagg
     1021 gcgtcatgga gtgaatgtgg aatctcctcc tttgccctcg ggcccgccga gtgtggtggg
     1081 ccagcctgga atgcctgatc cagcccccag gcctcgagga ccaaaactga agtttactcc
     1141 cgaagaggac gctctactgg tggagttgaa ggaaaacaag aacttgacgt ggaagcaaat
     1201 tgcagacttc ttcccgggcc gaacgagcgg taccttgcaa gtccgatact gcaccaagct
     1261 gaaggctaag gatgtagctt ggagtgacga aatggttcga tttgctcctg atgtatttct
     1321 acgcctgtct cacacatgct aatgaaggga ataggtacaa aggctgcagc gggcaatgca
     1381 cgagtacgag aacgatcggt ggcgcatcat tgcagggaag gttggaaatg gcttcacccc
     1441 agctgcttgc cgcgagaaag ccatgcagct ccatgagtaa aagcgttggg gaattttcat
     1501 atttatatct actgtcgcca gattcggccc tgcttggacc ctctgatctc cttactctcc
     1561 atattggttc aaatgtcggg tcaccgatag ggctggtggt gcaggcttgt tgtaggcacg
     1621 ggaggatgat cagcataact ctgagtcact atagggacgg gttgatgtaa ggtattaagt
     1681 gatgtatgat aattcatttt agcccggggg aacatatggc gccggcattt gttcgttcgc
     1741 aatgaaccga cactagcgtc cgctctcgca gtttagcacc ggctgatccc gggctgaacg
     1801 cggccattgc tcggccgggg catgtgttcc ttatctacgg cagaccgcag atgaccactg
     1861 gagcagatta tagaccctaa gccctaagcc ggacacccaa tcgagtaggt ctgcggacca
     1921 ggtcactgcg ggcagccgga gaagctccgc aaccaatcaa tccccggcgc tgactaaggg
     1981 caggcgacca cgggccgaag cggcttcaaa ctcacctcaa cctccaaact ccctcatctc
     2041 caaacgtcct tgccttgtct gccgtcattg caacccaccc accaggacac atgagcagca
     2101 agtcccagct cacctacact gcccgtgcca gcaagcatcc caatgctctg gcgaagaggc
     2161 tgttcgagat tgccgaggcc aagaaaacca atgtgactgt ctcggctgac gttaccacca
     2221 ctaaggagct actagatctt gctgaccgtc tcggtcccta cattgccgtg atcaaaaccc
     2281 acatcgatat cctctctgat ttcagcaacg agactattga gggacttaag gctctcgcgc
     2341 agaagcacaa ctttctcatc ttcgaggacc gcaagttcat tgacatcggc aacacggtcc
     2401 agaagcaata ccacggcggt accctccgta tctcggaatg ggcccacatc atcaactgca
     2461 gcattctccc tggtgagggt atcgtcgagg ctctcgctca gacggcgtct gcaccggact
     2521 tcgcctacgg ccccgaacgc ggtctgttga tcttggcaga gatgacctct aagggctcct
     2581 tggctaccgg ccagtacact acttcctcgg tcgattatgc ccggaaatac aagaacttcg
     2641 ttatgggatt cgtgtcgacg cgcgcgttgg gtgaggtgca gtcggaagtc agctctcctt
     2701 cggatgagga ggactttgtg gttttcacga ctggtgtgaa cattaacctc caagtcgcaa
     2761 ttgacctacg cttatgctgc agacgcgtta cgtatcggat ccagaattcg tgatatctga
     2821 attcgtcgac aagcttattt tttgtatact gttttgtgat agcacgaagt ttttccacgg
     2881 tatcttgtta aaaatatata tttgtggcgg gcttacctac atcaaattaa taagagacta
     2941 attataaact aaacacacaa gcaagctact ttagggtaaa agtttataaa tgcttttgac
     3001 gtataaacgt tgcttgtatt tattattaca attaaaggtg gatagaaaac ctagagacta
     3061 gttagaaact aatctcaggt ttgcgttaaa ctaaatcaga gcccgagagg ttaacagaac
     3121 ctagaagggg actagatatc cgggtaggga aacaaaaaaa aaaaacaaga cagccacata
     3181 ttagggagac tagttagaag ctagttccag gactaggaaa ataaaagaca atgataccac
     3241 agtctagttg acaactagat agattctaga ttgaggccaa agtctctgag atccaggtta
     3301 gttgcaacta atactagtta gtatctagtc tcctataact ctgaagctag aataacttac
     3361 tactattatc ctcaccactg ttcagctgcg caaacggagt gattgcaagg tgttcagaga
     3421 ctagttattg actagtcagt gactagcaat aactaacaag gtattaacct accatgtctg
     3481 ccatcaccct gcacttcctc gggctcagca gccttttcct cctcattttc atgctcattt
     3541 tccttgttta agactgtgac tagtcaaaga ctagtccaga accacaaagg agaaatgtct
     3601 taccactttc ttcattgctt gtctcttttg cattatccat gtctgcaact agttagagtc
     3661 tagttagtga ctagtccgac gaggacttgc ttgtctccgg attgttggag gaactctcca
     3721 gggcctcaag atccacaaca gagccttcta gatgactggt caataactag ttggtctttg
     3781 tctgagtctg actgacttac gaggttgcat actcgctccc tttgcctcgt caatcgatga
     3841 gaaaaagcgc caaaactcgc aatatggctt tgaaccacac ggtgctgaga ctagttagaa
     3901 tctagtccca aactagcttg gatagcttac ctttgccctt tgcgttgcga caggtcttgc
     3961 agggtatggt tcctttctca ccagctgatt tagctgcctt gctaccctca cggcggatct
     4021 gcataaagag tggctagagg ttataaatta gcactgatcc taggtacggg gctgaatgta
     4081 acttgccttt cctttctcat cgcgcggcaa gacaggcttg ctcaaattcc taccagtcac
     4141 aggggtatgc acggcgtacg gaccacttga actagtcaca gattagttag caactagtct
     4201 gcattgaatg gctgtactta cgggccctcg ccattgtcct gatcatttcc agcttcaccc
     4261 tcgttgctgc aaagtagtta gtgactagtc aaggactagt tgaaatggga gaagaaactc
     4321 acgaattctc gacaccctta gtattgtggt ccttggactt ggtgctgcta tatattagct
     4381 aatacactag ttagactcac agaaacttac gcagctcgct tgcgcttctt ggtaggagtc
     4441 ggggttggga gaacagtgcc ttcaaacaag ccttcatacc atgctacttg actagtcagg
     4501 gactagtcac caagtaatct agataggact tgcctttggc ctccatcagt tccttcatag
     4561 tgggaggtcc attgtgcaat gtaaactcca tgccgtggga gttcttgtcc ttcaagtgct
     4621 tgaccaatat gtttctgttg gcagagggaa cctgtcaact agttaataac tagtcagaaa
     4681 ctagtatagc agtagactca ctgtacgctt gaggcatccc ttcactcggc agtagacttc
     4741 atatggatgg atatcaggca cgccattgtc gtcctgtgga ctagtcagta actaggctta
     4801 aagctagtcg ggtcggctta ctatcttgaa atccggcagc gtaagctccc cgtccttaac
     4861 tgcctcgaga tagtgacagt actctgggga ctttcggaga tcgttatcgc gaatgctcgg
     4921 catactaatc gttgactagt cttggactag tcccgagcaa aaaggattgg aggaggagga
     4981 ggaaggtgag agtgagacaa agagcgaaat aagagcttca aaggctatct ctaagcagta
     5041 tgaaggttaa gtatctagtt cttgactaga tttaaaagag atttcgacta gttatgtacc
     5101 tggagtttgg atataggaat gtgttgtggt aacgaaatgt aagggggagg aaagaaaaag
     5161 tcggtcaaga ggtaactcta agtcggccat tcctttttgg gaggcgctaa ccataaacgg
     5221 catggtcgac ttagagttag ctcagggaat ttagggagtt atctgcgacc accgaggaac
     5281 ggcggaatgc caaagaatcc cgatggagct ctagctggcg gttgacaacc ccaccttttg
     5341 gcgtttctgc ggcgttgcag gcgggactgg atacttcgta gaaccagaaa ggcaaggcag
     5401 aacgcgctca gcaagagtgt tggaagtgat agcatgatgt gccttgttaa ctaggtcaaa
     5461 atctgcagta tgcttgatgt tatccaaagt gtgagagagg aaggtccaaa catacacgat
     5521 tgggagaggg cctaggtata agagtttttg agtagaacgc atgtgagccc agccatctcg
     5581 aggagattaa acacgggccg gcatttgatg gctatgttag taccccaatg gaaagcctga
     5641 gagtccagtg gcgctccggg gagaccaccg gtgccgctac agggcgcgtc aggtggcact
     5701 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5761 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5821 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5881 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5941 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6001 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6061 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6121 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6181 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6241 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6301 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6361 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6421 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6481 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggctct
     6541 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6601 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6661 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6721 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6781 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgtagatc tcatgaccaa
     6841 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6901 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6961 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7021 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
//
