LOCUS       sequence-181730-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 with split pyrG marker for homologous integration with linker DE
            (for 3 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: pMST1214_BB2_E_DE_pcoxA:pyrGtrunc_AMA1_2.8
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5062..5166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5167..5235,5236..6027)
                     /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(5385..5404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6253..6841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6742..6761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6995..7012
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacact
       61 agtcgcaaaa aaccccgccc ctgacagggc ggggtttttt cgcggtctcc agctggagac
      121 gggcacaagg ttcatgaacc gaagaggatg ggaaccccga ccatggcctg cggccgcagc
      181 tatcagtcct ctcagctgac gcccctcacg aaccctcgca ctctcccgaa atgctcttcg
      241 gtgtccatgg cgcatcatct cagcagcatc ccatgtcaaa ccatggcttt gggcccacgg
      301 actctgtggc cctgccgcaa catcaccatc accatcgact cccgccccat gcagcgctgc
      361 gcgccccagg gcgtcatgga gtgaatgtgg aatctcctcc tttgccctcg ggcccgccga
      421 gtgtggtggg ccagcctgga atgcctgatc cagcccccag gcctcgagga ccaaaactga
      481 agtttactcc cgaagaggac gctctactgg tggagttgaa ggaaaacaag aacttgacgt
      541 ggaagcaaat tgcagacttc ttcccgggcc gaacgagcgg taccttgcaa gtccgatact
      601 gcaccaagct gaaggctaag gatgtagctt ggagtgacga aatggttcga tttgctcctg
      661 atgtatttct acgcctgtct cacacatgct aatgaaggga ataggtacaa aggctgcagc
      721 gggcaatgca cgagtacgag aacgatcggt ggcgcatcat tgcagggaag gttggaaatg
      781 gcttcacccc agctgcttgc cgcgagaaag ccatgcagct ccatgagtaa aagcgttggg
      841 gaattttcat atttatatct actgtcgcca gattcggccc tgcttggacc ctctgatctc
      901 cttactctcc atattggttc aaatgtcggg tcaccgatag ggctggtggt gcaggcttgt
      961 tgtaggcacg ggaggatgat cagcataact ctgagtcact atagggacgg gttgatgtaa
     1021 ggtattaagt gatgtatgat aattcatttt agcccggggg aacatatggc gccggcattt
     1081 gttcgttcgc aatgaaccga cactagcgtc cgctctcgca gtttagcacc ggctgatccc
     1141 gggctgaacg cggccattgc tcggccgggg catgtgttcc ttatctacgg cagaccgcag
     1201 atgaccactg gagcagatta tagaccctaa gccctaagcc ggacacccaa tcgagtaggt
     1261 ctgcggacca ggtcactgcg ggcagccgga gaagctccgc aaccaatcaa tccccggcgc
     1321 tgactaaggg caggcgacca cgggccgaag cggcttcaaa ctcacctcaa cctccaaact
     1381 ccctcatctc caaacgtcct tgccttgtct gccgtcattg caacccaccc accaggacac
     1441 atgagcagca agtcccagct cacctacact gcccgtgcca gcaagcatcc caatgctctg
     1501 gcgaagaggc tgttcgagat tgccgaggcc aagaaaacca atgtgactgt ctcggctgac
     1561 gttaccacca ctaaggagct actagatctt gctgaccgtc tcggtcccta cattgccgtg
     1621 atcaaaaccc acatcgatat cctctctgat ttcagcaacg agactattga gggacttaag
     1681 gctctcgcgc agaagcacaa ctttctcatc ttcgaggacc gcaagttcat tgacatcggc
     1741 aacacggtcc agaagcaata ccacggcggt accctccgta tctcggaatg ggcccacatc
     1801 atcaactgca gcattctccc tggtgagggt atcgtcgagg ctctcgctca gacggcgtct
     1861 gcaccggact tcgcctacgg ccccgaacgc ggtctgttga tcttggcaga gatgacctct
     1921 aagggctcct tggctaccgg ccagtacact acttcctcgg tcgattatgc ccggaaatac
     1981 aagaacttcg ttatgggatt cgtgtcgacg cgcgcgttgg gtgaggtgca gtcggaagtc
     2041 agctctcctt cggatgagga ggactttgtg gttttcacga ctggtgtgaa cattaacctc
     2101 caagtcgcaa ttgacctacg cttatgctgc agacgcgtta cgtatcggat ccagaattcg
     2161 tgatatctga attcgtcgac aagcttattt tttgtatact gttttgtgat agcacgaagt
     2221 ttttccacgg tatcttgtta aaaatatata tttgtggcgg gcttacctac atcaaattaa
     2281 taagagacta attataaact aaacacacaa gcaagctact ttagggtaaa agtttataaa
     2341 tgcttttgac gtataaacgt tgcttgtatt tattattaca attaaaggtg gatagaaaac
     2401 ctagagacta gttagaaact aatctcaggt ttgcgttaaa ctaaatcaga gcccgagagg
     2461 ttaacagaac ctagaagggg actagatatc cgggtaggga aacaaaaaaa aaaaacaaga
     2521 cagccacata ttagggagac tagttagaag ctagttccag gactaggaaa ataaaagaca
     2581 atgataccac agtctagttg acaactagat agattctaga ttgaggccaa agtctctgag
     2641 atccaggtta gttgcaacta atactagtta gtatctagtc tcctataact ctgaagctag
     2701 aataacttac tactattatc ctcaccactg ttcagctgcg caaacggagt gattgcaagg
     2761 tgttcagaga ctagttattg actagtcagt gactagcaat aactaacaag gtattaacct
     2821 accatgtctg ccatcaccct gcacttcctc gggctcagca gccttttcct cctcattttc
     2881 atgctcattt tccttgttta agactgtgac tagtcaaaga ctagtccaga accacaaagg
     2941 agaaatgtct taccactttc ttcattgctt gtctcttttg cattatccat gtctgcaact
     3001 agttagagtc tagttagtga ctagtccgac gaggacttgc ttgtctccgg attgttggag
     3061 gaactctcca gggcctcaag atccacaaca gagccttcta gatgactggt caataactag
     3121 ttggtctttg tctgagtctg acttacgagg ttgcatactc gctccctttg cctcgtcaat
     3181 cgatgagaaa aagcgccaaa actcgcaata tggctttgaa ccacacggtg ctgagactag
     3241 ttagaatcta gtcccaaact agcttggata gcttaccttt gccctttgcg ttgcgacagg
     3301 tcttgcaggg tatggttcct ttctcaccag ctgatttagc tgccttgcta ccctcacggc
     3361 ggatctgcat aaagagtggc tagaggttat aaattagcac tgatcctagg tacggggctg
     3421 aatgtaactt gcctttcctt tctcatcgcg cggcaagaca ggcttgctca aattcctacc
     3481 agtcacaggg gtatgcacgg cgtacggacc acttgaacta gtcacagatt agttagcaac
     3541 tagtctgcat tgaatggctg tacttacggg ccctcgccat tgtcctgatc atttccagct
     3601 tcaccctcgt tgctgcaaag tagttagtga ctagtcaagg actagttgaa atgggagaag
     3661 aaactcacga attctcgaca cccttagtat tgtggtcctt ggacttggtg ctgctatata
     3721 ttagctaata cactagttag actcacagaa acttacgcag ctcgcttgcg cttcttggta
     3781 ggagtcgggg ttgggagaac agtgccttca aacaagcctt cataccatgc tacttgacta
     3841 gtcagggact agtcaccaag taatctagat aggacttgcc tttggcctcc atcagttcct
     3901 tcatagtggg aggtccattg tgcaatgtaa actccatgcc gtgggagttc ttgtccttca
     3961 agtgcttgac caatatgttt ctgttggcag agggaacctg tcaactagtt aataactagt
     4021 cagaaactag tatagcagta gactcactgt acgcttgagg catcccttca ctcggcagta
     4081 gacttcatat ggatggatat caggcacgcc attgtcgtcc tgtggactag tcagtaacta
     4141 ggcttaaagc tagtcgggtc ggcttactat cttgaaatcc ggcagcgtaa gctccccgtc
     4201 cttaactgcc tcgagatagt gacagtactc tggggacttt cggagatcgt tatcgcgaat
     4261 gctcggcata ctaatcgttg actagtcttg gactagtccc gagcaaaaag gattggagga
     4321 ggaggaggaa ggtgagagtg agacaaagag cgaaataaga gcttcaaagg ctatctctaa
     4381 gcagtatgaa ggttaagtat ctagttcttg actagattta aaagagattt cgactagtta
     4441 tgtacctgga gtttggatat aggaatgtgt tgtggtaacg aaatgtaagg gggaggaaag
     4501 aaaaagtcgg tcaagaggta actctaagtc ggccattcct ttttgggagg cgctaaccat
     4561 aaacggcatg gtcgacttag agttagctca gggaatttag ggagttatct gcgaccaccg
     4621 aggaacggcg gaatgccaaa gaatcccgat ggagctctag ctggcggttg acaaccccac
     4681 cttttggcgt ttctgcggcg ttgcaggcgg gactggatac ttcgtagaac cagaaaggca
     4741 aggcagaacg cgctcagcaa gagtgttgga agtgatagca tgatgtgcct tgttaactag
     4801 gtcaaaatct gcagtatgct tgatgttatc caaagtgtga gagaggaagg tccaaacata
     4861 cacgattggg agagggccta ggtataagag tttttgagta gaacgcatgt gagcccagcc
     4921 atctcgagga gattaaacac gggccggcat ttgatggcta tgttagtacc ccaatggaaa
     4981 gcctgagagt ccagtggcgc tggagggaga ccaccggtgc cgctacaggg cgcgtcaggt
     5041 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5101 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5161 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5281 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5341 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5401 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5461 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5581 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5701 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5761 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5881 ggctctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5941 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6001 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6061 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6121 ctcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg tagatctcat
     6181 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6241 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6301 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6361 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6421 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6481 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6541 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6601 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6661 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6721 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6781 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6841 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6901 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6961 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     7021 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgatt
//
