LOCUS       sequence-254032-        5235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 3. CAD transcription unit with the TDH3 
            promoter and TDH1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5235)
  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 5235)
  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-bc-Ptdh3-CAD-Ttdh1
FEATURES             Location/Qualifiers
     source          1..5235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    144..177
                     /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)"
     promoter        554..841
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     711..734
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     protein_bind    complement(2686..2719)
                     /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(3189..3206)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3189..3205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3304..3321)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3475..4063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3555..3574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4185..4994)
                     /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(4995..5086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5153..5172
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga gtagggaact gccaggcatc
       61 aaataaaacg aaaggctcag tcgaaagact gggcctttcg cccgggctaa ttagggggtg
      121 tcgcccttat tcgactctat agtgaagttc ctattctcta gaaagtatag gaacttctga
      181 agtggggcgg atcgatgtac acaaccgact gcacccaaac gaacacaaat cttagcagtg
      241 cttagtcaaa aaattagcct tttaattctg ctgtaacccg tacatgccca aaataggggg
      301 cgggttacac agaatatata acatcgtagg tgtctgggtg aacagtttat tcctggcatc
      361 cactaaatat aatggagccc gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc
      421 accaaaatat tgttttcttc accaaccatc agttcatagg tccattctct tagcgcaact
      481 acagagaaca ggggcacaaa caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac
      541 ctgcctggag taaatgatga cacaaggcaa ttgacccacg catgtatcta tctcattttc
      601 ttacaccttc tattaccttc tgctctctct gatttggaaa aagctgaaaa aaaaggttga
      661 aaccagttcc ctgaaattat tcccctactt gactaataag tatataaaga cggtaggtat
      721 tgattgtaat tctgtaaatc tatttcttaa acttcttaaa ttctactttt atagttagtc
      781 ttttttttag ttttaaaaca ccaagaactt agtttcgaat aaacacacat aaacaaacaa
      841 aatgaccaag caatctgctg actccaatgc caagtctggt gtcacttccg aaatctgtca
      901 ctgggcttcc aacttggcta ctgacgacat tccatctgat gtcttggaaa gagccaagta
      961 cttgattttg gacggtattg cctgtgcttg ggttggtgct cgtgttccat ggtctgaaaa
     1021 gtacgttcaa gctaccatgt ccttcgaacc tccaggtgct tgtcgtgtca tcggttacgg
     1081 tcaaaaattg ggtccagttg ctgctgccat gaccaactct gcctttattc aagccactga
     1141 attggatgac taccactctg aagctccatt gcactctgct tccattgttc taccagctgt
     1201 tttcgctgct tctgaagtct tggctgaaca aggtaagacc atctctggta tcgatgttat
     1261 cttagctgcc attgtcggtt tcgaatctgg tccaagaatc ggtaaggcca tctacggttc
     1321 tgacttgttg aacaacggtt ggcattgtgg tgccgtttac ggtgctccag ctggtgcttt
     1381 ggctaccggt aagttgttgg gtttgactcc agactccatg gaagatgctt tgggtatcgc
     1441 ttgtacccaa gcttgtggtt tgatgtctgc tcaatacggt ggtatggtta agagagttca
     1501 acatggtttc gctgccagaa acggtctatt aggtggtttg ttggctcacg gtggttacga
     1561 agctatgaag ggtgtcttgg aaagatctta cggtggtttc ttgaagatgt tcaccaaggg
     1621 taacggtaga gaacctccat acaaggaaga agaagttgtt gccggtttag gttctttctg
     1681 gcacactttc accatcagaa tcaaattgta cgcttgttgt ggtttagtcc acggtccagt
     1741 tgaagccatt gaaaacttac aaggtcgtta cccagaattg ttgaacagag ctaacttgtc
     1801 caacatcaga cacgttcacg ttcaattatc cactgcttcc aactctcact gtggttggat
     1861 tccagaagaa agaccaatct cctccattgc tggtcaaatg tctgttgctt acattttggc
     1921 tgtccaattg gttgaccaac aatgtttgtt gtctcaattc tccgaattcg atgacaactt
     1981 ggaaagacca gaagtctggg atttggctag aaaggtcacc tcttctcaat ctgaagaatt
     2041 tgaccaagat ggtaactgtt tgtctgctgg tagagtcaga attgaattca acgacggttc
     2101 ttccatcact gaatccgttg aaaagccatt aggtgtcaag gaaccaatgc caaacgaaag
     2161 aatcttgcac aaatacagaa ctttggctgg ttccgtcact gacgaatcca gagtcaagga
     2221 aatcgaagat ttggttttgg gtttggacag attgaccgat atctctccat tattggaatt
     2281 gttgaactgt ccagtcaaat ctccattggg tatcaagtaa aataaagcaa tcttgatgag
     2341 gataatgatt tttttttgaa tatacataaa tactaccgtt tttctgctag attttgtgaa
     2401 gacgtaaata agtacatatt actttttaag ccaagacaag attaagcatt aactttaccc
     2461 ttttctcttc taagtttcaa tactagttat cactgtttaa aagttatggc gagaacgtcg
     2521 gcggttaaaa tatattaccc tgaacgtggt gaattgaagt tctaggatgg tttaaagatt
     2581 tttccttttt gggaaataag taaacaatat attgctgcct tcctcacgct ttccggcatc
     2641 ttccagacca cagtatatcc atccgcctcc tgttgaaaat gaagtgaagt tcctatactt
     2701 tctagagaat aggaacttct atagtgagtc gaataagggc gacacaaaat ttattctaaa
     2761 tgcataataa atactgataa catcttatag tttgtattat attttgtatt atcgttgaca
     2821 tgtataattt tgatatcaaa aactgatttt ccctttatta ttttcgagat ttattttctt
     2881 aattctcttt aacaaactag aaatattgta tatacaaaaa atcataaata atagatgaat
     2941 agtttaatta taggtgttca tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc
     3001 ttttttctca tttataaggt taaataattc tcatatatca agcaaagtga caggcgccct
     3061 taaatattct gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg
     3121 ggtttttacg ttatttgcgg attaacgatt actcgttatc agaaccgccc agggggcccg
     3181 agcttaagac tggccgtcgt tttacaacac agaaagagtt tgtagaaacg caaaaaggcc
     3241 atccgtcagg ggccttctgc ttagtttgat gcctggcagt tccctactct cgccttccgc
     3301 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3361 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3421 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3481 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3541 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3601 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3661 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3721 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3781 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3841 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt gggctaacta
     3901 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3961 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4021 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4081 ttctacgggg tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg
     4141 gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct ctgcttagaa aaactcatcg
     4201 agcatcaaat gaaattgcaa tttattcata tcaggattat caataccata tttttgaaaa
     4261 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     4321 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     4381 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     4441 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     4501 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgaggcga
     4561 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgagtgcaa ccggcgcagg
     4621 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     4681 aacgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     4741 aaatgcttga tggtcggaag tggcataaat tccgtcagcc agtttagtct gaccatctca
     4801 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     4861 ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     4921 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga cgtttcccgt
     4981 tgaatatggc tcatattctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5041 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt
     5101 acaaccaatt aaccaattct gaacattatc gcgagcccat ttatacctga atatggctca
     5161 taacacccct tgtttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     5221 ctcagaagtg aaacg
//
