LOCUS       sequence-254140-        4690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 6. CSC transcription unit with the S. bayanus
            TDH3 promoter and EFM1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4690)
  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 4690)
  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-ef-Psbtdh3-CSC-Tefm1
FEATURES             Location/Qualifiers
     source          1..4690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    172..205
                     /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)"
     protein_bind    complement(2169..2202)
                     /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(2672..2689)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2672..2688)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2787..2804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2958..3546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3038..3057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3668..4477)
                     /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(4478..4569)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4636..4655
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggactcccc
       61 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      121 gcctttcgcc cgggctaatt agggggtgtc gcccttattc gactctatag tgaagttcct
      181 attctctaga aagtatagga acttctgaag tggggaaata accacaaaca tccttcccat
      241 atgctcggtc gtgcttgttg tacctgtgcc attcatcttt cacctgccat tagtaacccg
      301 acttctcatt gagcgggtta cggcagccac aggccacatt ccgaatgtct gggtgagcgg
      361 tcccttttcc agcatccact aaatatctcg gatcccgctt tttaatctgg cttcctgaaa
      421 aaaatcaatg gagtgatgca aactgactgg agcaaaaagc tgacacaagg caatcgacct
      481 acgtgtctgt ctattttctc acaccttcta ttaccttcta actctctggg ttggaaaaaa
      541 ctgaaaaaaa ggttgtctcc agtttccaca aatcatcccc ctgtttgatt aataaatata
      601 taaagacgac aactatcgat cataaactca taaaactata actcctttac acttcttatt
      661 ttatagttat tctattttaa ttcttattga ttttaaaacc ccaagaactt agtttcgaaa
      721 acacacacac acaaacaatt aaaaatggct gacaccaagg ccaagttgac cttgaacggt
      781 gacactgctg tcgaattgga tgttttgaaa ggtactttgg gtcaagatgt cattgatatc
      841 agaactttgg gttccaaggg tgttttcacc ttcgacccag gtttcacctc tactgcttct
      901 tgtgaatcca agatcacttt catcgatggt gacgaaggta tcctattaca cagaggtttc
      961 ccaattgacc aattagctac tgactccaac tacttggaag tttgttacat cttgttgaat
     1021 ggtgaaaagc caactcaaga acaatacgac gaatttaaaa ccaccgttac cagacacacc
     1081 atgattcacg aacaaatcac cagattattc cacgctttcc gtcgtgactc ccacccaatg
     1141 gctgtcatgt gtggtatcac tggtgctttg gctgctttct accatgactc tttggatgtc
     1201 aacaacccaa gacacagaga aattgccgct ttcttgttgt tgtccaagat gccaaccatg
     1261 gctgctatgt gttacaagta ctccatcggt caacctttcg tttacccaag aaacgatttg
     1321 tcttacgccg gtaacttctt gaacatgatg ttctccactc catgtgaacc ttacgaagtt
     1381 aacccaattt tggaaagagc catggacaga atcttgatct tgcacgctga ccatgaacaa
     1441 aacgcttcta cttctactgt tagaactgcc ggttcttctg gtgctaaccc atttgcttgt
     1501 atcgctgctg gtattgcttc tttatggggt ccagctcatg gtggtgccaa cgaagctgct
     1561 ttgaagatgt tggaagaaat ttcttctgtc aagcacattc cagaatttgt cagaagagct
     1621 aaggacaaga acgactcttt cagattgatg ggtttcggtc accgtgtcta caagaactac
     1681 gacccaagag ctaccgtcat gagagaaacc tgtcacgaag ttttgaagga attgggtacc
     1741 aaggatgact tgttggaagt tgccatggaa ttggaaaaca ttgctttgaa cgacccatac
     1801 ttcatcgaaa agaaattgta cccaaacgtc gatttctact ccggtatcat cttaaaggct
     1861 atgggtattc catcttccat gttcaccgtt atctttgcta tggccagaac tgttggttgg
     1921 atcgctcact ggtccgaaat gcactctgat ggtatgaaga ttgccagacc aagacaatta
     1981 tacactggtt acgaaaagag agatttcaaa tctgatatca agagataaat ttgatctgta
     2041 gcctaagtat aaaattctac gtatgtatat atttacatgc aattttttct ttttccaatt
     2101 catgcctcga aaccttcgaa tccagccagc atgtcgacac ccacaagatg tagtgcacaa
     2161 aatgaagtga agttcctata ctttctagag aataggaact tctatagtga gtcgaataag
     2221 ggcgacacaa aatttattct aaatgcataa taaatactga taacatctta tagtttgtat
     2281 tatattttgt attatcgttg acatgtataa ttttgatatc aaaaactgat tttcccttta
     2341 ttattttcga gatttatttt cttaattctc tttaacaaac tagaaatatt gtatatacaa
     2401 aaaatcataa ataatagatg aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac
     2461 gtatcttatt taaagtgcgt tgcttttttc tcatttataa ggttaaataa ttctcatata
     2521 tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat gctctttccc taaactcccc
     2581 ccataaaaaa acccgccgaa gcgggttttt acgttatttg cggattaacg attactcgtt
     2641 atcagaaccg cccagggggc ccgagcttaa gactggccgt cgttttacaa cacagaaaga
     2701 gtttgtagaa acgcaaaaag gccatccgtc aggggccttc tgcttagttt gatgcctggc
     2761 agttccctac tctcgccttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3361 ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc
     3601 gtaactcacg ttaagggatt ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat
     3661 gctctgctta gaaaaactca tcgagcatca aatgaaattg caatttattc atatcaggat
     3721 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     3781 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     3841 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     3901 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     3961 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     4021 gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     4081 gaatcgagtg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     4141 caggatattc ttctaatacc tggaacgctg tttttccggg gatcgcagtg gtgagtaacc
     4201 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagtggcata aattccgtca
     4261 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     4321 tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc gcacctgatt
     4381 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     4441 atcgcggcct cgacgtttcc cgttgaatat ggctcatatt cttccttttt caatattatt
     4501 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4561 ataaacaaat aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc
     4621 catttatacc tgaatatggc tcataacacc ccttgtttgc ctggcggcag tagcgcggtg
     4681 gtcccacctg
//
