LOCUS       sequence-244779-        4974 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 4974)
  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 4974)
  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..4974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(810..901)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     968..987
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1194..1227
                     /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)"
     terminator      3109..3296
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3164..3188)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(3473..3506)
                     /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(3976..3993)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3976..3992)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4091..4108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4262..4850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4342..4361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4974,1..809))
                     /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"
ORIGIN
        1 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
       61 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
      121 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
      181 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
      241 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
      301 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
      361 gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgag
      421 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
      481 tcttctaata cctggaacgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
      541 tcaggagtac ggataaaatg cttgatggtc ggaagtggca taaattccgt cagccagttt
      601 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
      661 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
      721 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
      781 ctcgacgttt cccgttgaat atggctcata ttcttccttt ttcaatatta ttgaagcatt
      841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
      901 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
      961 cctgaatatg gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc
     1021 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc
     1081 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1141 gggcctttcg cccgggctaa ttagggggtg tcgcccttat tcgactctat agtgaagttc
     1201 ctattctcta gaaagtatag gaacttctga agtggggaag cgacttccaa tcgctttgca
     1261 tatccagtac cacacccaca ggcgtttgtg cgtgcaacat atatacacaa ttacagtaac
     1321 aataacaaga ggacagatac taccaaaatg tgtggggaag cgggtaagct gccacagcaa
     1381 ttaatgcaca acatttaacc tacattcttc cttatcggat cctcaaaacc cttaaaaaca
     1441 tatgcctcac cctaacatat tttccaatta accctcaata tttctctgtc acccggcctc
     1501 tattttccat tttcttcttt acccgccacg cgtttttttc tttcaaattt ttttcttctt
     1561 tcttcttttt cttccacgtc ctcttgcata aataaataaa ccgttttgaa accaaactcg
     1621 cctctctctc tcctttttga aatatttttg ggtttgtttg atcctttcct tcccaatctc
     1681 tcttgtttaa tatatattca tttatatcac gctctctttt tatcttcctt tttttcctct
     1741 ctcttgtatt cttccttccc ctttctactc aaaccaagaa gaaaaagaaa aggtcaatct
     1801 ttgttaaaga ataggatctt ctactacatc agcttttaga tttttcacgc ttactgcttt
     1861 tttcttccca agatcgaaaa tttactgaat taacaatggg tcacggtgac actgaatctc
     1921 caaacccaac caccaccact gaaggttctg gtcaaaacga acctgaaaag aagggtcgtg
     1981 acattccatt atggagaaag tgtgttatca ctttcgttgt ttcctggatg actttggttg
     2041 tcactttctc ctccacctgt ttgttgccag ctgctccaga aattgctaac gaattcgata
     2101 tgaccgtcga aaccattaac atttccaacg ctggtgtttt ggttgccatg ggttactctt
     2161 ctttgatctg gggtccaatg aacaaattgg ttggtagaag aacctcttac aacttggcca
     2221 tctccatgtt gtgtgcctgt tctgctggta ctgctgctgc catcaacgaa gaaatgttca
     2281 ttgctttccg tgtcttgtct ggcttgaccg gtacttcttt catggtttcc ggtcaaaccg
     2341 tcttggctga tatctttgaa ccagtttaca gaggtactgc tgtcggtttc ttcatggctg
     2401 gtactctatc cggtccagcc attggtccat gtgtcggtgg tgtcattgtc actttcacct
     2461 cctggagagt tatcttctgg ttacaattgg gtatgtctgg tttaggtttg gttttgtctc
     2521 tattattctt cccaaagatc gaaggtaact ctgaaaaggt ttctactgct ttcaagccaa
     2581 ccactttggt caccatcatc tccaagttct ctccaaccga tgtcttgaag caatgggttt
     2641 acccaaatgt ctttttggct gatttgtgtt gtggtttgtt ggccatcact caatactcca
     2701 tcttgacttc tgccagagct atcttcaact ccagattcca tttgaccacc gctttggttt
     2761 ccggtttatt ctacttggct ccaggtgctg gtttcttgat tggttctttg gttggtggta
     2821 aattgtctga cagaaccgtc agaagataca ttgtcaagag aggtttcaga ttacctcaag
     2881 acagattgca ctctggtttg atcactttgt ttgctgtctt gccagctggt actttgatct
     2941 acggttggac tttgcaagag gacaagggtg acatggttgt tccaatcatt gctgctttct
     3001 ttgctggttg gggtttgatg ggttctttca actgtttgaa cacctacgtt gctggtttat
     3061 tccacacttt gatctacttg ttcccattgt gtacctgtcc acaataaagc gaatttctta
     3121 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     3181 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     3241 aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg
     3301 ccgagcaaat gcctgcaaat cgctccccat ttcacccaat tgtagatatg ctaactccag
     3361 caatgagttg atgaatctcg gtgtgtattt tatgtcctca gaggacaacc tcttgcccat
     3421 cgaacgtaca agtactcctc tgttctctcc ttcctttgct ttaaaatgaa gtgaagttcc
     3481 tatactttct agagaatagg aacttctata gtgagtcgaa taagggcgac acaaaattta
     3541 ttctaaatgc ataataaata ctgataacat cttatagttt gtattatatt ttgtattatc
     3601 gttgacatgt ataattttga tatcaaaaac tgattttccc tttattattt tcgagattta
     3661 ttttcttaat tctctttaac aaactagaaa tattgtatat acaaaaaatc ataaataata
     3721 gatgaatagt ttaattatag gtgttcatca atcgaaaaag caacgtatct tatttaaagt
     3781 gcgttgcttt tttctcattt ataaggttaa ataattctca tatatcaagc aaagtgacag
     3841 gcgcccttaa atattctgac aaatgctctt tccctaaact ccccccataa aaaaacccgc
     3901 cgaagcgggt ttttacgtta tttgcggatt aacgattact cgttatcaga accgcccagg
     3961 gggcccgagc ttaagactgg ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa
     4021 aaaggccatc cgtcaggggc cttctgctta gtttgatgcc tggcagttcc ctactctcgc
     4081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggg
     4681 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4861 tgatcttttc tacggggtct gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg
     4921 gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg cttt
//
