LOCUS       sequence-254121-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 6. CSC transcription unit with the TEF1 
            promoter and PDC1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5040)
  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 5040)
  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-Ptef1-CSC-Tpdc1
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    73..106
                     /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        344..757
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     protein_bind    complement(2420..2453)
                     /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(2923..2940)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2923..2939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3038..3055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3209..3797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3289..3308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3919..4728)
                     /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(4729..4820)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4887..4906
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aaggctcagt cgaaagactg ggcctttcgc ccgggctaat tagggggtgt cgcccttatt
       61 cgactctata gtgaagttcc tattctctag aaagtatagg aacttctgaa gtggggaaat
      121 aaccacaaac atccttccca tatgctcggt cgtgcttgtt gtacctgtgc ttggctgata
      181 atagcgtata aacaatgcat actttgtacg ttcaaaatac aatgcagtag atatatttat
      241 gcatattaca tataatacat atcacatagg aagcaacagg cgcgttggac ttttaatttt
      301 cgaggaccgc gaatccttac atcacaccca atcccccaca agtgatcccc cacacaccat
      361 agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc
      421 gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta
      481 gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagacacc gcctcgtttc
      541 tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt
      601 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
      661 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
      721 tgctcattag aaagaaagca tagcaatcta atctaagttt taattacaaa atggctgaca
      781 ccaaggccaa gttgaccttg aacggtgaca ctgctgtcga attggatgtt ttgaaaggta
      841 ctttgggtca agatgtcatt gatatcagaa ctttgggttc caagggtgtt ttcaccttcg
      901 acccaggttt cacctctact gcttcttgtg aatccaagat cactttcatc gatggtgacg
      961 aaggtatcct attacacaga ggtttcccaa ttgaccaatt agctactgac tccaactact
     1021 tggaagtttg ttacatcttg ttgaatggtg aaaagccaac tcaagaacaa tacgacgaat
     1081 ttaaaaccac cgttaccaga cacaccatga ttcacgaaca aatcaccaga ttattccacg
     1141 ctttccgtcg tgactcccac ccaatggctg tcatgtgtgg tatcactggt gctttggctg
     1201 ctttctacca tgactctttg gatgtcaaca acccaagaca cagagaaatt gccgctttct
     1261 tgttgttgtc caagatgcca accatggctg ctatgtgtta caagtactcc atcggtcaac
     1321 ctttcgttta cccaagaaac gatttgtctt acgccggtaa cttcttgaac atgatgttct
     1381 ccactccatg tgaaccttac gaagttaacc caattttgga aagagccatg gacagaatct
     1441 tgatcttgca cgctgaccat gaacaaaacg cttctacttc tactgttaga actgccggtt
     1501 cttctggtgc taacccattt gcttgtatcg ctgctggtat tgcttcttta tggggtccag
     1561 ctcatggtgg tgccaacgaa gctgctttga agatgttgga agaaatttct tctgtcaagc
     1621 acattccaga atttgtcaga agagctaagg acaagaacga ctctttcaga ttgatgggtt
     1681 tcggtcaccg tgtctacaag aactacgacc caagagctac cgtcatgaga gaaacctgtc
     1741 acgaagtttt gaaggaattg ggtaccaagg atgacttgtt ggaagttgcc atggaattgg
     1801 aaaacattgc tttgaacgac ccatacttca tcgaaaagaa attgtaccca aacgtcgatt
     1861 tctactccgg tatcatctta aaggctatgg gtattccatc ttccatgttc accgttatct
     1921 ttgctatggc cagaactgtt ggttggatcg ctcactggtc cgaaatgcac tctgatggta
     1981 tgaagattgc cagaccaaga caattataca ctggttacga aaagagagat ttcaaatctg
     2041 atatcaagag ataaagcgat ttaatctcta attattagtt aaagttttat aagcattttt
     2101 atgtaacgaa aaataaattg gttcatatta ttactgcact gtcacttacc atggaaagac
     2161 cagacaagaa gttgccgaca gtctgttgaa ttggcctggt taggcttaag tctgggtccg
     2221 cttctttaca aatttggaga atttctctta aacgatatgt atattctttt cgttggaaaa
     2281 gatgtcttcc aaaaaaaaaa ccgatgaatt agtggaacca aggaaaaaaa aagaggtatc
     2341 cttgattaag gaacacctcg aaaccttcga atccagccag catgtcgaca cccacaagat
     2401 gtagtgcaca aaatgaagtg aagttcctat actttctaga gaataggaac ttctatagtg
     2461 agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt
     2521 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
     2581 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
     2641 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     2701 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     2761 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     2821 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     2881 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     2941 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     3001 tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3601 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3841 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     3901 gtcagcgtaa tgctctgctt agaaaaactc atcgagcatc aaatgaaatt gcaatttatt
     3961 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     4021 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     4081 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     4141 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     4201 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     4261 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
     4321 attacaaaca ggaatcgagt gcaaccggcg caggaacact gccagcgcat caacaatatt
     4381 ttcacctgaa tcaggatatt cttctaatac ctggaacgct gtttttccgg ggatcgcagt
     4441 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagtggcat
     4501 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4561 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     4621 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4681 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcatat tcttcctttt
     4741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4801 tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat
     4861 tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgtttg cctggcggca
     4921 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     4981 atggtagtgt ggggactccc catgcgagag tagggaactg ccaggcatca aataaaacga
//
