LOCUS       sequence-253028-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 1. ITE transcription unit with the ACT1 
            promoter and ADH1 terminator. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  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 4968)
  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-5a-Pact1-ITE-Tadh1
FEATURES             Location/Qualifiers
     source          1..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1641..1828
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1696..1720)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(2005..2038)
                     /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(2508..2525)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2508..2524)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2623..2640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2794..3382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2874..2893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3504..4313)
                     /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(4314..4405)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4472..4491
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4698..4731
                     /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 taaccctcaa tatttctctg tcacccggcc tctattttcc attttcttct ttacccgcca
       61 cgcgtttttt tctttcaaat ttttttcttc tttcttcttt ttcttccacg tcctcttgca
      121 taaataaata aaccgttttg aaaccaaact cgcctctctc tctccttttt gaaatatttt
      181 tgggtttgtt tgatcctttc cttcccaatc tctcttgttt aatatatatt catttatatc
      241 acgctctctt tttatcttcc tttttttcct ctctcttgta ttcttccttc ccctttctac
      301 tcaaaccaag aagaaaaaga aaaggtcaat ctttgttaaa gaataggatc ttctactaca
      361 tcagctttta gatttttcac gcttactgct tttttcttcc caagatcgaa aatttactga
      421 attaacaatg ggtcacggtg acactgaatc tccaaaccca accaccacca ctgaaggttc
      481 tggtcaaaac gaacctgaaa agaagggtcg tgacattcca ttatggagaa agtgtgttat
      541 cactttcgtt gtttcctgga tgactttggt tgtcactttc tcctccacct gtttgttgcc
      601 agctgctcca gaaattgcta acgaattcga tatgaccgtc gaaaccatta acatttccaa
      661 cgctggtgtt ttggttgcca tgggttactc ttctttgatc tggggtccaa tgaacaaatt
      721 ggttggtaga agaacctctt acaacttggc catctccatg ttgtgtgcct gttctgctgg
      781 tactgctgct gccatcaacg aagaaatgtt cattgctttc cgtgtcttgt ctggcttgac
      841 cggtacttct ttcatggttt ccggtcaaac cgtcttggct gatatctttg aaccagttta
      901 cagaggtact gctgtcggtt tcttcatggc tggtactcta tccggtccag ccattggtcc
      961 atgtgtcggt ggtgtcattg tcactttcac ctcctggaga gttatcttct ggttacaatt
     1021 gggtatgtct ggtttaggtt tggttttgtc tctattattc ttcccaaaga tcgaaggtaa
     1081 ctctgaaaag gtttctactg ctttcaagcc aaccactttg gtcaccatca tctccaagtt
     1141 ctctccaacc gatgtcttga agcaatgggt ttacccaaat gtctttttgg ctgatttgtg
     1201 ttgtggtttg ttggccatca ctcaatactc catcttgact tctgccagag ctatcttcaa
     1261 ctccagattc catttgacca ccgctttggt ttccggttta ttctacttgg ctccaggtgc
     1321 tggtttcttg attggttctt tggttggtgg taaattgtct gacagaaccg tcagaagata
     1381 cattgtcaag agaggtttca gattacctca agacagattg cactctggtt tgatcacttt
     1441 gtttgctgtc ttgccagctg gtactttgat ctacggttgg actttgcaag aggacaaggg
     1501 tgacatggtt gttccaatca ttgctgcttt ctttgctggt tggggtttga tgggttcttt
     1561 caactgtttg aacacctacg ttgctggttt attccacact ttgatctact tgttcccatt
     1621 gtgtacctgt ccacaataaa gcgaatttct tatgatttat gatttttatt attaaataag
     1681 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1741 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     1801 tcgctcttat tgaccacacc tctaccggca tgccgagcaa atgcctgcaa atcgctcccc
     1861 atttcaccca attgtagata tgctaactcc agcaatgagt tgatgaatct cggtgtgtat
     1921 tttatgtcct cagaggacaa cctcttgccc atcgaacgta caagtactcc tctgttctct
     1981 ccttcctttg ctttaaaatg aagtgaagtt cctatacttt ctagagaata ggaacttcta
     2041 tagtgagtcg aataagggcg acacaaaatt tattctaaat gcataataaa tactgataac
     2101 atcttatagt ttgtattata ttttgtatta tcgttgacat gtataatttt gatatcaaaa
     2161 actgattttc cctttattat tttcgagatt tattttctta attctcttta acaaactaga
     2221 aatattgtat atacaaaaaa tcataaataa tagatgaata gtttaattat aggtgttcat
     2281 caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct tttttctcat ttataaggtt
     2341 aaataattct catatatcaa gcaaagtgac aggcgccctt aaatattctg acaaatgctc
     2401 tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga
     2461 ttaacgatta ctcgttatca gaaccgccca gggggcccga gcttaagact ggccgtcgtt
     2521 ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct
     2581 tagtttgatg cctggcagtt ccctactctc gccttccgct tcctcgctca ctgactcgct
     2641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3181 aggcggtgct acagagttct tgaagtggtg ggctaactac ggctacacta gaagaacagt
     3241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3421 gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg tcatgagctt gcgccgtccc
     3481 gtcaagtcag cgtaatgctc tgcttagaaa aactcatcga gcatcaaatg aaattgcaat
     3541 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     3601 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     3661 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     3721 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     3781 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     3841 aaaccgttat tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa
     3901 ggacaattac aaacaggaat cgagtgcaac cggcgcagga acactgccag cgcatcaaca
     3961 atattttcac ctgaatcagg atattcttct aatacctgga acgctgtttt tccggggatc
     4021 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaagt
     4081 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     4141 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caagcgatag
     4201 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     4261 tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct catattcttc
     4321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4381 gaatgtattt agaaaaataa acaaataggg gtcagtgtta caaccaatta accaattctg
     4441 aacattatcg cgagcccatt tatacctgaa tatggctcat aacacccctt gtttgcctgg
     4501 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     4561 cgccgatggt agtgtgggga ctccccatgc gagagtaggg aactgccagg catcaaataa
     4621 aacgaaaggc tcagtcgaaa gactgggcct ttcgcccggg ctaattaggg ggtgtcgccc
     4681 ttattcgact ctatagtgaa gttcctattc tctagaaagt ataggaactt ctgaagtggg
     4741 gaagcgactt ccaatcgctt tgcatatcca gtaccacacc cacaggcgtt tgtgcaacat
     4801 atatacacaa ttacagtaac aataacaaga ggacagatac taccaaaatg tgtggggaag
     4861 cgggtaagct gccacagcaa ttaatgcaca acatttaacc tacattcttc cttatcggat
     4921 cctcaaaacc cttaaaaaca tatgcctcac cctaacatat tttccaat
//
