LOCUS       sequence-254147-        5032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 7. ACDH transcription unit with the VMA6 
            promoter and RPS14A terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5032)
  AUTHORS   Young EM, Zhao Z, Gielesen BEM, Wu L, Benjamin Gordon D, Roubos JA, 
            Voigt CA
  TITLE     Iterative algorithm-guided design of massive strain libraries, 
            applied to itaconic acid production in yeast.
  JOURNAL   Metab Eng. 2018 Jul;48:33-43. doi: 10.1016/j.ymben.2018.05.002. Epub
            2018 May 9.
  PUBMED    29753070
REFERENCE   2  (bases 1 to 5032)
  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: "Metab 
            Eng."; date: "2018-07"; volume: "48:33-43. doi"; pages: " 
            10.1016/j.ymben.2018.05.002. Epub 2018 May 9"
COMMENT     Sequence Label: pDSM-f3-Pvma6-ACDH-Trps14a
FEATURES             Location/Qualifiers
     source          1..5032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2249..2282)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(2752..2769)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2752..2768)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2867..2884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3038..3626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3118..3137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3748..4557)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(4558..4649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4716..4735
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4942..4975
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 acgtgcaagg caagaaaaaa aattatgacc catacatttt ggtgcatggg tgatattttg
       61 tctttattat tattagtaac tttcagatta agacaaggaa gaaagaagga aaaaaggtag
      121 gacatgaagc atgagagata atcaaatttg ctgaatgctg tgatactggt atacaaatag
      181 aagtgctgaa gttcaagtta tttggtaaca tgcaagtttc attaatattt tgttattatg
      241 tatttcaagg acaaaatgaa ttgtaggaag atgaaaaagt gatttcatcc cccaacgatt
      301 catactctgg gccgtaccct tcttatctta cagacgcaaa gtcagcagat tcttgtatta
      361 tggaactaag taatttccgc gcacctattg ttacgctgat atctgcgtac aaagtgacgt
      421 atttttaaaa tatcttcgat atactgcaaa aacaaaagaa caattgacct cttaaccctt
      481 taaggtatgg cataaagaac gctaagagct agcaaaagag tataccattc ggatcgtgtt
      541 gctaaagtct tgcgaaaatg gaatctttgg aattggaaca attagtcaag aaggttttgt
      601 tggaaaaatt ggctgaacaa aaggaagttc caaccaagac caccacccaa ggtgccaagt
      661 ccggtgtttt cgacaccgtc gatgaagctg tccaagctgc tgtcattgct caaaactgtt
      721 acaaggaaaa atctttggaa gaaagaagaa acgttgtcaa ggccatcaga gaagctttgt
      781 acccagaaat cgaaaccatt gccaccagag ctgttgctga aaccggtatg ggtaatgtca
      841 ctgacaagat cttgaagaac actttggcca tcgaaaagac cccaggtgtt gaagatttgt
      901 acactgaagt tgccactggt gacaacggta tgactttgta cgaattgtct ccatacggtg
      961 tcatcggtgc tgttgcccca tctaccaacc caactgaaac tttgatctgt aactccattg
     1021 gtatgttggc tgctggtaat gctgttttct actctcctca cccaggtgcc aagaacatct
     1081 ctttatggtt aatcgaaaaa ttgaacacca ttgtccgtga ctcttgtggt atcgacaact
     1141 tgattgtcac tgttgccaag ccttccatcc aagctgctca agaaatgatg aaccatccaa
     1201 aggtcccatt gttggttatc actggtggtc caggtgttgt cttgcaagct atgcaatctg
     1261 gtaagaaggt cattggtgct ggtgctggta acccaccatc tatcgtcgat gaaactgcta
     1321 acattgaaaa ggctgccgct gatatcgttg acggtgcttc tttcgaccac aacatcctat
     1381 gtattgctga aaaatccgtt gttgccgttg actccattgc tgatttctta ttattccaaa
     1441 tggaaaagaa cggtgctttg cacgttacca acccatctga tatccaaaaa ttggaaaagg
     1501 ttgctgtcac tgacaagggt gtcaccaaca agaaattggt tggtaagtct gctactgaaa
     1561 tcttgaagga agctggtatt gcttgtgact tcactccaag attaatcatt gtcgaaactg
     1621 aaaagtccca cccatttgcc accgttgaat tgttgatgcc aattgtccca gttgtcagag
     1681 ttccagactt cgatgaagct ttggaagttg ccatcgaatt ggaacaaggt ttgcaccaca
     1741 ctgctaccat gcactctcaa aacatctcca gattgaacaa ggctgctaga gacatgcaaa
     1801 cttccatctt tgtcaagaac ggtccatctt tcgctggttt aggtttcaga ggtgaaggtt
     1861 ccactacttt caccattgct accccaactg gtgaaggtac caccaccgct agacatttcg
     1921 ctagaagaag aagatgtgtt ttgactgatg gtttctccat acgttaaatt atgcatgtat
     1981 tgtacttgta ttgccgtatt attttttaca gttaaaaaat gtgtacatat aattatatag
     2041 cgcccataat caaatcagct catacgtcaa tttagtaata aaaaaaagcc cttataacct
     2101 tttagttaag aagattcaag attgcgattt gattaacgtc attacggaaa tgtaaggaca
     2161 caattaccaa gagttacaaa atcgcctcag aaagcctgta tgcgaagcca caatcctttc
     2221 caacagacca tactaagtaa aatgaagtga agttcctata ctttctagag aataggaact
     2281 tctatagtga gtcgaataag ggcgacacaa aatttattct aaatgcataa taaatactga
     2341 taacatctta tagtttgtat tatattttgt attatcgttg acatgtataa ttttgatatc
     2401 aaaaactgat tttcccttta ttattttcga gatttatttt cttaattctc tttaacaaac
     2461 tagaaatatt gtatatacaa aaaatcataa ataatagatg aatagtttaa ttataggtgt
     2521 tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt tgcttttttc tcatttataa
     2581 ggttaaataa ttctcatata tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat
     2641 gctctttccc taaactcccc ccataaaaaa acccgccgaa gcgggttttt acgttatttg
     2701 cggattaacg attactcgtt atcagaaccg cccagggggc ccgagcttaa gactggccgt
     2761 cgttttacaa cacagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggggccttc
     2821 tgcttagttt gatgcctggc agttccctac tctcgccttc cgcttcctcg ctcactgact
     2881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2941 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3001 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3061 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3421 atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa
     3481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3661 ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcttgcgccg
     3721 tcccgtcaag tcagcgtaat gctctgctta gaaaaactca tcgagcatca aatgaaattg
     3781 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     3841 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     3901 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     3961 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     4021 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     4081 aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt
     4141 aaaaggacaa ttacaaacag gaatcgagtg caaccggcgc aggaacactg ccagcgcatc
     4201 aacaatattt tcacctgaat caggatattc ttctaatacc tggaacgctg tttttccggg
     4261 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     4321 aagtggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     4381 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg
     4441 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     4501 agcatccatg ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcatatt
     4561 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4621 atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca attaaccaat
     4681 tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc ccttgtttgc
     4741 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     4801 gtagcgccga tggtagtgtg gggactcccc atgcgagagt agggaactgc caggcatcaa
     4861 ataaaacgaa aggctcagtc gaaagactgg gcctttcgcc cgggctaatt agggggtgtc
     4921 gcccttattc gactctatag tgaagttcct attctctaga aagtatagga acttctgaag
     4981 tggggaaacc ttcgaatcca gccagcatgt cgacacccac aagatgtagt gc
//
