LOCUS       sequence-254089-        5747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 4. ACO transcription unit with the TEF1 
            promoter and YOL036w terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5747)
  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 5747)
  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-cd-Ptef1-ACO-Tyol036w
FEATURES             Location/Qualifiers
     source          1..5747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    126..159
                     /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        223..622
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     protein_bind    complement(3180..3213)
                     /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(3683..3700)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3683..3699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3798..3815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3969..4557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4049..4068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4679..5488)
                     /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(5489..5580)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5647..5666
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgtggggact ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
       61 agtcgaaaga ctgggccttt cgcccgggct aattaggggg tgtcgccctt attcgactct
      121 atagtgaagt tcctattctc tagaaagtat aggaacttct gaagtgggga cgctttccgg
      181 catcttccag accacagtat atccatccgc ctcctgttgg tgcatagctt caaaatgttt
      241 ctactccttt tttactcttc cagattttct cggacaccgc gcatcgccgt accacttcaa
      301 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
      361 cccgtactaa aggtttggaa aagaaaaaag tgaccgcctc gtttcttttt cttcgtcgaa
      421 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt tttgattttt
      481 ttctctttcg atgacctccc attgatattt aagttaataa acggtgttca atttctcaag
      541 tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc attagaaaga
      601 aagcatagca atctaatcta agttttaatt acaaaatgac tgtttccaac ttgaccagag
      661 actccaaggt taaccaaaac ttgttggaag atcattcttt catcaactac aagcaaaatg
      721 tcgaaacttt ggatatcgtc agaaagagat tgaacagacc attcacctac gctgaaaaga
      781 ttttgtacgg tcacttggat gacccacacg gtcaagatat ccaaagaggt gtctcctact
      841 tgaaactaag accagatcgt gttgcttgtc aagatgctac tgctcaaatg gctatcttac
      901 aattcatgtc cgctggtttg cctcaagttg ccaagccagt caccgtccac tgtgaccatt
      961 tgatccaagc tcaagtcggt ggtgaaaagg acttgaagag agccattgac ttgaacaagg
     1021 aagtctacga cttcttggct tctgccactg ctaaatacaa catgggtttc tggaagccag
     1081 gttccggtat catccaccaa atcgttttgg aaaactatgc cttcccaggt gctttgatca
     1141 tcggtactga ctcccacact ccaaatgccg gtggtctagg tcaattggcc atcggtgttg
     1201 gtggtgctga tgctgttgac gtcatggctg gtagaccatg ggaattgaag gctccaaaga
     1261 ttttgggtgt taagttgacc ggtaagatga acggttggac ttctccaaag gacatcatct
     1321 tgaaattggc tggtatcact actgttaagg gtggtactgg taagattgtc gaatactttg
     1381 gtgacggtgt cgacactttc tctgctaccg gtatgggtac catctgtaac atgggtgctg
     1441 aaattggtgc caccacttct gttttcccat tcaacaaatc catgattgaa tacttggaag
     1501 ctaccggtag aggtaagatt gctgatttcg ctaagttata ccacaaggac ttgttgtctg
     1561 ccgacaagga cgctgaatac gatgaagttg tcgaaattga cttgaacact ttggaaccat
     1621 acatcaacgg tccattcacc ccagatttgg ctaccccagt ttctaagatg aaggaagttg
     1681 ccgttgctaa caactggcca ttagatgtta gagttggttt gattggttct tgtaccaact
     1741 cctcttacga agatatgtcc agatctgctt ccattgtcaa ggatgctgct gctcacggtt
     1801 tgaaatctaa gaccatcttc actgttaccc caggttctga acaaatcaga gccaccatcg
     1861 aacgtgacgg tcaattggaa actttcaagg aatttggtgg tattgtcttg gctaacgctt
     1921 gtggtccatg tattggtcaa tgggacagaa gagatatcaa gaagggtgac aagaacacca
     1981 tcgtttcctc ttacaacaga aacttcactt ctagaaacga tggtaaccca caaacccacg
     2041 cctttgttgc ttctccagaa ttagtcactg ctttcgctat tgctggtgac ttgagattca
     2101 acccattaac cgacaaattg aaggacaagg acggtaacga atttatgttg aagcctcctc
     2161 atggtgatgg tttaccacaa agaggttacg atgctggtga aaacacctac caagctccac
     2221 cagccgacag atccaccgtc gaagtcaagg tttctccaac ttctgacaga ttacaattgt
     2281 tgaaaccttt caagccatgg gatggtaagg acgctaagga catgccaatc ttaatcaagg
     2341 ctgttggtaa gactaccacc gaccacattt ccatggctgg tccatggttg aaatacagag
     2401 gtcacttgga aaacatctcc aacaactaca tgattggtgc cattaatgcc gaaaacaaga
     2461 aggctaactg tgtcaagaac gtttacactg gtgaatacaa gggtgttcca gacactgcca
     2521 gagactacag agatcaaggt atcaaatggg ttgtcatcgg tgacgaaaac ttcggtgaag
     2581 gttcttctcg tgaacacgct gctttggaac caagattctt gggtggtttc gctattatta
     2641 ccaaatcttt cgctcgtatt cacgaaacca acttgaagaa gcaaggtcta ttgccattga
     2701 acttcaagaa cccagccgac tacgacaaga tcaacccaga tgacagaatt gacatcttag
     2761 gtttggctga attggctcca ggtaagccag tcaccatgag agttcaccca aagaacggta
     2821 agccatggga tgctgtcttg actcacactt tcaacgatga acaaatcgaa tggttcaaat
     2881 acggttctgc tttgaacaag atcaaggctg atgaaaagaa gtaaaggaaa aaggtcttgg
     2941 ctatatataa cggcagacaa aatataagta tacacgtata tatggtggta aacacgcata
     3001 tactgtatgc catgtattta ccattacata gttatttacg cactctataa aaagttaaca
     3061 ttgcatttta ataaattcct taaattactc taattaggat ggtagcccta cctttcctca
     3121 acgttgtcca ggtttgtatc cacgtgtgtc cgttccgcca atattccgca aaatgaagtg
     3181 aagttcctat actttctaga gaataggaac ttctatagtg agtcgaataa gggcgacaca
     3241 aaatttattc taaatgcata ataaatactg ataacatctt atagtttgta ttatattttg
     3301 tattatcgtt gacatgtata attttgatat caaaaactga ttttcccttt attattttcg
     3361 agatttattt tcttaattct ctttaacaaa ctagaaatat tgtatataca aaaaatcata
     3421 aataatagat gaatagttta attataggtg ttcatcaatc gaaaaagcaa cgtatcttat
     3481 ttaaagtgcg ttgctttttt ctcatttata aggttaaata attctcatat atcaagcaaa
     3541 gtgacaggcg cccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa
     3601 aacccgccga agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc
     3661 gcccaggggg cccgagctta agactggccg tcgttttaca acacagaaag agtttgtaga
     3721 aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt tgatgcctgg cagttcccta
     3781 ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3841 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3901 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3961 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4021 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4081 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4141 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4201 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4261 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4321 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4381 tggtgggcta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4441 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4501 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4561 gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgcg cgtaactcac
     4621 gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgctt
     4681 agaaaaactc atcgagcatc aaatgaaatt gcaatttatt catatcagga ttatcaatac
     4741 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     4801 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     4861 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     4921 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     4981 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     5041 cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgagt
     5101 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     5161 cttctaatac ctggaacgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat
     5221 caggagtacg gataaaatgc ttgatggtcg gaagtggcat aaattccgtc agccagttta
     5281 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     5341 actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat
     5401 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     5461 tcgacgtttc ccgttgaata tggctcatat tcttcctttt tcaatattat tgaagcattt
     5521 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5581 taggggtcag tgttacaacc aattaaccaa ttctgaacat tatcgcgagc ccatttatac
     5641 ctgaatatgg ctcataacac cccttgtttg cctggcggca gtagcgcggt ggtcccacct
     5701 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtag
//
