LOCUS       sequence-176368-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 with split pyrG marker for homologous integration with linker BC
            (for 1 expression cassette).
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: pMST1212_BB2_E_BC_pcoxA:pyrGtrunc_AMA1_2.8
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     169..188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     422..439
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5559..5663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5664..5732,5733..6524)
                     /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(5882..5901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6750..7070,1..268)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
       61 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      361 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      421 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      481 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      541 tgagcgcaac actagtcgca aaaaaccccg cccctgacag ggcggggttt tttcgcggtc
      601 tccccgggga gacgggcaca aggttcatga accgaagagg atgggaaccc cgaccatggc
      661 ctgcggccgc agctatcagt cctctcagct gacgcccctc acgaaccctc gcactctccc
      721 gaaatgctct tcggtgtcca tggcgcatca tctcagcagc atcccatgtc aaaccatggc
      781 tttgggccca cggactctgt ggccctgccg caacatcacc atcaccatcg actcccgccc
      841 catgcagcgc tgcgcgcccc agggcgtcat ggagtgaatg tggaatctcc tcctttgccc
      901 tcgggcccgc cgagtgtggt gggccagcct ggaatgcctg atccagcccc caggcctcga
      961 ggaccaaaac tgaagtttac tcccgaagag gacgctctac tggtggagtt gaaggaaaac
     1021 aagaacttga cgtggaagca aattgcagac ttcttcccgg gccgaacgag cggtaccttg
     1081 caagtccgat actgcaccaa gctgaaggct aaggatgtag cttggagtga cgaaatggtt
     1141 cgatttgctc ctgatgtatt tctacgcctg tctcacacat gctaatgaag ggaataggta
     1201 caaaggctgc agcgggcaat gcacgagtac gagaacgatc ggtggcgcat cattgcaggg
     1261 aaggttggaa atggcttcac cccagctgct tgccgcgaga aagccatgca gctccatgag
     1321 taaaagcgtt ggggaatttt catatttata tctactgtcg ccagattcgg ccctgcttgg
     1381 accctctgat ctccttactc tccatattgg ttcaaatgtc gggtcaccga tagggctggt
     1441 ggtgcaggct tgttgtaggc acgggaggat gatcagcata actctgagtc actataggga
     1501 cgggttgatg taaggtatta agtgatgtat gataattcat tttagcccgg gggaacatat
     1561 ggcgccggca tttgttcgtt cgcaatgaac cgacactagc gtccgctctc gcagtttagc
     1621 accggctgat cccgggctga acgcggccat tgctcggccg gggcatgtgt tccttatcta
     1681 cggcagaccg cagatgacca ctggagcaga ttatagaccc taagccctaa gccggacacc
     1741 caatcgagta ggtctgcgga ccaggtcact gcgggcagcc ggagaagctc cgcaaccaat
     1801 caatccccgg cgctgactaa gggcaggcga ccacgggccg aagcggcttc aaactcacct
     1861 caacctccaa actccctcat ctccaaacgt ccttgccttg tctgccgtca ttgcaaccca
     1921 cccaccagga cacatgagca gcaagtccca gctcacctac actgcccgtg ccagcaagca
     1981 tcccaatgct ctggcgaaga ggctgttcga gattgccgag gccaagaaaa ccaatgtgac
     2041 tgtctcggct gacgttacca ccactaagga gctactagat cttgctgacc gtctcggtcc
     2101 ctacattgcc gtgatcaaaa cccacatcga tatcctctct gatttcagca acgagactat
     2161 tgagggactt aaggctctcg cgcagaagca caactttctc atcttcgagg accgcaagtt
     2221 cattgacatc ggcaacacgg tccagaagca ataccacggc ggtaccctcc gtatctcgga
     2281 atgggcccac atcatcaact gcagcattct ccctggtgag ggtatcgtcg aggctctcgc
     2341 tcagacggcg tctgcaccgg acttcgccta cggccccgaa cgcggtctgt tgatcttggc
     2401 agagatgacc tctaagggct ccttggctac cggccagtac actacttcct cggtcgatta
     2461 tgcccggaaa tacaagaact tcgttatggg attcgtgtcg acgcgcgcgt tgggtgaggt
     2521 gcagtcggaa gtcagctctc cttcggatga ggaggacttt gtggttttca cgactggtgt
     2581 gaacattaac ctccaagtcg caattgacct acgcttatgc tgcagacgcg ttacgtatcg
     2641 gatccagaat tcgtgatatc tgaattcgtc gacaagctta ttttttgtat actgttttgt
     2701 gatagcacga agtttttcca cggtatcttg ttaaaaatat atatttgtgg cgggcttacc
     2761 tacatcaaat taataagaga ctaattataa actaaacaca caagcaagct actttagggt
     2821 aaaagtttat aaatgctttt gacgtataaa cgttgcttgt atttattatt acaattaaag
     2881 gtggatagaa aacctagaga ctagttagaa actaatctca ggtttgcgtt aaactaaatc
     2941 agagcccgag aggttaacag aacctagaag gggactagat atccgggtag ggaaacaaaa
     3001 aaaaaaaaca agacagccac atattaggga gactagttag aagctagttc caggactagg
     3061 aaaataaaag acaatgatac cacagtctag ttgacaacta gatagattct agattgaggc
     3121 caaagtctct gagatccagg ttagttgcaa ctaatactag ttagtatcta gtctcctata
     3181 actctgaagc tagaataact tactactatt atcctcacca ctgttcagct gcgcaaacgg
     3241 agtgattgca aggtgttcag agactagtta ttgactagtc agtgactagc aataactaac
     3301 aaggtattaa cctaccatgt ctgccatcac cctgcacttc ctcgggctca gcagcctttt
     3361 cctcctcatt ttcatgctca ttttccttgt ttaagactgt gactagtcaa agactagtcc
     3421 agaaccacaa aggagaaatg tcttaccact ttcttcattg cttgtctctt ttgcattatc
     3481 catgtctgca actagttaga gtctagttag tgactagtcc gacgaggact tgcttgtctc
     3541 cggattgttg gaggaactct ccagggcctc aagatccaca acagagcctt ctagatgact
     3601 ggtcaataac tagttggtct ttgtctgagt ctgactgact tacgaggttg catactcgct
     3661 ccctttgcct cgtcaatcga tgagaaaaag cgccaaaact cgcaatatgg ctttgaacca
     3721 cacggtgctg agactagtta gaatctagtc ccaaactagc ttggatagct tacctttgcc
     3781 ctttgcgttg cgacaggtct tgcagggtat ggttcctttc tcaccagctg atttagctgc
     3841 cttgctaccc tcacggcgga tctgcataaa gagtggctag aggttataaa ttagcactga
     3901 tcctaggtac ggggctgaat gtaacttgcc tttcctttct catcgcgcgg caagacaggc
     3961 ttgctcaaat tcctaccagt cacaggggta tgcacggcgt acggaccact tgaactagtc
     4021 acagattagt tagcaactag tctgcattga atggctgtac ttacgggccc tcgccattgt
     4081 cctgatcatt tccagcttca ccctcgttgc tgcaaagtag ttagtgacta gtcaaggact
     4141 agttgaaatg ggagaagaaa ctcacgaatt ctcgacaccc ttagtattgt ggtccttgga
     4201 cttggtgctg ctatatatta gctaatacac tagttagact cacagaaact tacgcagctc
     4261 gcttgcgctt cttggtagga gtcggggttg ggagaacagt gccttcaaac aagccttcat
     4321 accatgctac ttgactagtc agggactagt caccaagtaa tctagatagg acttgccttt
     4381 ggcctccatc agttccttca tagtgggagg tccattgtgc aatgtaaact ccatgccgtg
     4441 ggagttcttg tccttcaagt gcttgaccaa tatgtttctg ttggcagagg gaacctgtca
     4501 actagttaat aactagtcag aaactagtat agcagtagac tcactgtacg cttgaggcat
     4561 cccttcactc ggcagtagac ttcatatgga tggatatcag gcacgccatt gtcgtcctgt
     4621 ggactagtca gtaactaggc ttaaagctag tcgggtcggc ttactatctt gaaatccggc
     4681 agcgtaagct ccccgtcctt aactgcctcg agatagtgac agtactctgg ggactttcgg
     4741 agatcgttat cgcgaatgct cggcatacta atcgttgact agtcttggac tagtcccgag
     4801 caaaaaggat tggaggagga ggaggaaggt gagagtgaga caaagagcga aataagagct
     4861 tcaaaggcta tctctaagca gtatgaaggt taagtatcta gttcttgact agatttaaaa
     4921 gagatttcga ctagttatgt acctggagtt tggatatagg aatgtgttgt ggtaacgaaa
     4981 tgtaaggggg aggaaagaaa aagtcggtca agaggtaact ctaagtcggc cattcctttt
     5041 tgggaggcgc taaccataaa cggcatggtc gacttagagt tagctcaggg aatttaggga
     5101 gttatctgcg accaccgagg aacggcggaa tgccaaagaa tcccgatgga gctctagctg
     5161 gcggttgaca accccacctt ttggcgtttc tgcggcgttg caggcgggac tggatacttc
     5221 gtagaaccag aaaggcaagg cagaacgcgc tcagcaagag tgttggaagt gatagcatga
     5281 tgtgccttgt taactaggtc aaaatctgca gtatgcttga tgttatccaa agtgtgagag
     5341 aggaaggtcc aaacatacac gattgggaga gggcctaggt ataagagttt ttgagtagaa
     5401 cgcatgtgag cccagccatc tcgaggagat taaacacggg ccggcatttg atggctatgt
     5461 tagtacccca atggaaagcc tgagagtcca gtggcgctaa ttggagacca ccggtgccgc
     5521 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5581 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5641 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5701 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5761 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5821 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5881 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5941 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6001 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6061 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6121 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6181 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6241 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6301 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6361 ctggagccgg tgagcgtggc tctcgcggta tcattgcagc actggggcca gatggtaagc
     6421 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6481 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6541 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6601 agatcctttt tgataatctc atgaacaata aaactgtctg cttacataaa cagtaataca
     6661 aggggtgtag atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6721 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6781 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6841 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     6901 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6961 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7021 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
//
