LOCUS       sequence-254062-        5680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 4. ACO transcription unit with the YKT6 
            promoter and PRM9 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  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 5680)
  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-Pykt6-ACO-Tprm9
FEATURES             Location/Qualifiers
     source          1..5680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    163..196
                     /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)"
     protein_bind    complement(3150..3183)
                     /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(3653..3670)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3653..3669)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3768..3785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3939..4527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4019..4038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4649..5458)
                     /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(5459..5550)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5617..5636
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggactccc catgcgagag
       61 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgc
      121 ccgggctaat tagggggtgt cgcccttatt cgactctata gtgaagttcc tattctctag
      181 aaagtatagg aacttctgaa gtggggacgc tttccggcat cttccagacc acagtatatc
      241 catccgcctc ctgttggtgc tttactgata atatacactc tttggatcga gcccacttcc
      301 agttggtaat tggtgttcca caatttcagc attacatgtt tttaaaccaa aattcggctc
      361 cttttccctt tttttcttat tgggtggcgt gccgtacaga acgattggct tggtgtgaaa
      421 tcaagagcaa gcacaataga tatcaacatg aacaatatac aaaagtctct ggcacagttt
      481 gactgcgtta gaccaggcta gggcatttct gaagctttac gtatcactag agaagttatt
      541 ttggcaatga ctgtttccaa cttgaccaga gactccaagg ttaaccaaaa cttgttggaa
      601 gatcattctt tcatcaacta caagcaaaat gtcgaaactt tggatatcgt cagaaagaga
      661 ttgaacagac cattcaccta cgctgaaaag attttgtacg gtcacttgga tgacccacac
      721 ggtcaagata tccaaagagg tgtctcctac ttgaaactaa gaccagatcg tgttgcttgt
      781 caagatgcta ctgctcaaat ggctatctta caattcatgt ccgctggttt gcctcaagtt
      841 gccaagccag tcaccgtcca ctgtgaccat ttgatccaag ctcaagtcgg tggtgaaaag
      901 gacttgaaga gagccattga cttgaacaag gaagtctacg acttcttggc ttctgccact
      961 gctaaataca acatgggttt ctggaagcca ggttccggta tcatccacca aatcgttttg
     1021 gaaaactatg ccttcccagg tgctttgatc atcggtactg actcccacac tccaaatgcc
     1081 ggtggtctag gtcaattggc catcggtgtt ggtggtgctg atgctgttga cgtcatggct
     1141 ggtagaccat gggaattgaa ggctccaaag attttgggtg ttaagttgac cggtaagatg
     1201 aacggttgga cttctccaaa ggacatcatc ttgaaattgg ctggtatcac tactgttaag
     1261 ggtggtactg gtaagattgt cgaatacttt ggtgacggtg tcgacacttt ctctgctacc
     1321 ggtatgggta ccatctgtaa catgggtgct gaaattggtg ccaccacttc tgttttccca
     1381 ttcaacaaat ccatgattga atacttggaa gctaccggta gaggtaagat tgctgatttc
     1441 gctaagttat accacaagga cttgttgtct gccgacaagg acgctgaata cgatgaagtt
     1501 gtcgaaattg acttgaacac tttggaacca tacatcaacg gtccattcac cccagatttg
     1561 gctaccccag tttctaagat gaaggaagtt gccgttgcta acaactggcc attagatgtt
     1621 agagttggtt tgattggttc ttgtaccaac tcctcttacg aagatatgtc cagatctgct
     1681 tccattgtca aggatgctgc tgctcacggt ttgaaatcta agaccatctt cactgttacc
     1741 ccaggttctg aacaaatcag agccaccatc gaacgtgacg gtcaattgga aactttcaag
     1801 gaatttggtg gtattgtctt ggctaacgct tgtggtccat gtattggtca atgggacaga
     1861 agagatatca agaagggtga caagaacacc atcgtttcct cttacaacag aaacttcact
     1921 tctagaaacg atggtaaccc acaaacccac gcctttgttg cttctccaga attagtcact
     1981 gctttcgcta ttgctggtga cttgagattc aacccattaa ccgacaaatt gaaggacaag
     2041 gacggtaacg aatttatgtt gaagcctcct catggtgatg gtttaccaca aagaggttac
     2101 gatgctggtg aaaacaccta ccaagctcca ccagccgaca gatccaccgt cgaagtcaag
     2161 gtttctccaa cttctgacag attacaattg ttgaaacctt tcaagccatg ggatggtaag
     2221 gacgctaagg acatgccaat cttaatcaag gctgttggta agactaccac cgaccacatt
     2281 tccatggctg gtccatggtt gaaatacaga ggtcacttgg aaaacatctc caacaactac
     2341 atgattggtg ccattaatgc cgaaaacaag aaggctaact gtgtcaagaa cgtttacact
     2401 ggtgaataca agggtgttcc agacactgcc agagactaca gagatcaagg tatcaaatgg
     2461 gttgtcatcg gtgacgaaaa cttcggtgaa ggttcttctc gtgaacacgc tgctttggaa
     2521 ccaagattct tgggtggttt cgctattatt accaaatctt tcgctcgtat tcacgaaacc
     2581 aacttgaaga agcaaggtct attgccattg aacttcaaga acccagccga ctacgacaag
     2641 atcaacccag atgacagaat tgacatctta ggtttggctg aattggctcc aggtaagcca
     2701 gtcaccatga gagttcaccc aaagaacggt aagccatggg atgctgtctt gactcacact
     2761 ttcaacgatg aacaaatcga atggttcaaa tacggttctg ctttgaacaa gatcaaggct
     2821 gatgaaaaga agtaaacaga tgacgggaga cactagcaca caactttacc aggcaaggta
     2881 tttgacgcta gcatgtgtcc aattcagtgt catttatgat tttttgtagt aggatataaa
     2941 tatatacagc gctccaaata gtgcggttgc cccaaaaaca ccacggaacc tcatctgttc
     3001 tcgtactttg ttgtgacaaa gtagctcact gccttattat cacattttca ttatgcaacg
     3061 cttcggaaaa tacgatgttg aaaatcctca acgttgtcca ggtttgtatc cacgtgtgtc
     3121 cgttccgcca atattccgca aaatgaagtg aagttcctat actttctaga gaataggaac
     3181 ttctatagtg agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg
     3241 ataacatctt atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat
     3301 caaaaactga ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa
     3361 ctagaaatat tgtatataca aaaaatcata aataatagat gaatagttta attataggtg
     3421 ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata
     3481 aggttaaata attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa
     3541 tgctctttcc ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt
     3601 gcggattaac gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg
     3661 tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt
     3721 ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac
     3781 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4321 tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga
     4381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4561 gctcagtgga acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc
     4621 gtcccgtcaa gtcagcgtaa tgctctgctt agaaaaactc atcgagcatc aaatgaaatt
     4681 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     4741 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     4801 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     4861 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     4921 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     4981 caaccaaacc gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt
     5041 taaaaggaca attacaaaca ggaatcgagt gcaaccggcg caggaacact gccagcgcat
     5101 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaacgct gtttttccgg
     5161 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     5221 gaagtggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     5281 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc
     5341 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     5401 cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcatat
     5461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5521 tatttgaatg tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa
     5581 ttctgaacat tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgtttg
     5641 cctggcggca gtagcgcggt ggtcccacct gaccccatgc
//
