LOCUS       sequence-181724-        7066 bp    DNA     circular SYN 15-APR-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 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 Apr 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..7066
                     /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        5555..5659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5660..5728,5729..6520)
                     /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(5878..5897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6746..7066,1..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 ctgacttacg aggttgcata ctcgctccct
     3661 ttgcctcgtc aatcgatgag aaaaagcgcc aaaactcgca atatggcttt gaaccacacg
     3721 gtgctgagac tagttagaat ctagtcccaa actagcttgg atagcttacc tttgcccttt
     3781 gcgttgcgac aggtcttgca gggtatggtt cctttctcac cagctgattt agctgccttg
     3841 ctaccctcac ggcggatctg cataaagagt ggctagaggt tataaattag cactgatcct
     3901 aggtacgggg ctgaatgtaa cttgcctttc ctttctcatc gcgcggcaag acaggcttgc
     3961 tcaaattcct accagtcaca ggggtatgca cggcgtacgg accacttgaa ctagtcacag
     4021 attagttagc aactagtctg cattgaatgg ctgtacttac gggccctcgc cattgtcctg
     4081 atcatttcca gcttcaccct cgttgctgca aagtagttag tgactagtca aggactagtt
     4141 gaaatgggag aagaaactca cgaattctcg acacccttag tattgtggtc cttggacttg
     4201 gtgctgctat atattagcta atacactagt tagactcaca gaaacttacg cagctcgctt
     4261 gcgcttcttg gtaggagtcg gggttgggag aacagtgcct tcaaacaagc cttcatacca
     4321 tgctacttga ctagtcaggg actagtcacc aagtaatcta gataggactt gcctttggcc
     4381 tccatcagtt ccttcatagt gggaggtcca ttgtgcaatg taaactccat gccgtgggag
     4441 ttcttgtcct tcaagtgctt gaccaatatg tttctgttgg cagagggaac ctgtcaacta
     4501 gttaataact agtcagaaac tagtatagca gtagactcac tgtacgcttg aggcatccct
     4561 tcactcggca gtagacttca tatggatgga tatcaggcac gccattgtcg tcctgtggac
     4621 tagtcagtaa ctaggcttaa agctagtcgg gtcggcttac tatcttgaaa tccggcagcg
     4681 taagctcccc gtccttaact gcctcgagat agtgacagta ctctggggac tttcggagat
     4741 cgttatcgcg aatgctcggc atactaatcg ttgactagtc ttggactagt cccgagcaaa
     4801 aaggattgga ggaggaggag gaaggtgaga gtgagacaaa gagcgaaata agagcttcaa
     4861 aggctatctc taagcagtat gaaggttaag tatctagttc ttgactagat ttaaaagaga
     4921 tttcgactag ttatgtacct ggagtttgga tataggaatg tgttgtggta acgaaatgta
     4981 agggggagga aagaaaaagt cggtcaagag gtaactctaa gtcggccatt cctttttggg
     5041 aggcgctaac cataaacggc atggtcgact tagagttagc tcagggaatt tagggagtta
     5101 tctgcgacca ccgaggaacg gcggaatgcc aaagaatccc gatggagctc tagctggcgg
     5161 ttgacaaccc caccttttgg cgtttctgcg gcgttgcagg cgggactgga tacttcgtag
     5221 aaccagaaag gcaaggcaga acgcgctcag caagagtgtt ggaagtgata gcatgatgtg
     5281 ccttgttaac taggtcaaaa tctgcagtat gcttgatgtt atccaaagtg tgagagagga
     5341 aggtccaaac atacacgatt gggagagggc ctaggtataa gagtttttga gtagaacgca
     5401 tgtgagccca gccatctcga ggagattaaa cacgggccgg catttgatgg ctatgttagt
     5461 accccaatgg aaagcctgag agtccagtgg cgctaattgg agaccaccgg tgccgctaca
     5521 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5581 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5641 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5761 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5821 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5941 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6061 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6241 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6361 agccggtgag cgtggctctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6541 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6601 cctttttgat aatctcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     6661 gtgtagatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6721 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6781 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6841 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6901 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6961 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7021 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaac
//
