LOCUS       sequence-254094-        7299 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast pathway position 5. PYC transcription unit with the PGK1 
            promoter and TPI1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  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 7299)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-de-Ppgk1-PYC-Ttpi1
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(4217..4250)
                     /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(4457..4476)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        4543..4634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4635..5444
                     /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"
     rep_origin      5566..6154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6055..6074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6308..6325
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6423..6440
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6424..6440
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6910..6943
                     /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 tattaatctt tatttctttt gagaaggtgg tagagtttct tcttccaaaa caaccaacaa
       61 gtcagaagca tcgacggatt caccgtcacg gataaagaca tctttgactt gaccatcagc
      121 tggagaggaa acaaccattt ccatcttcat agcagacaaa acggcgatag attcaccctt
      181 cttgaccaaa gaacccttgt gaaccttgac ttcaatgatg acaccagcca ttggagcacc
      241 gatttggtgg gtgtcgtgga catcagcctt tggcttggca acagattgaa tgttttgaga
      301 tttgtcagca actctgatct ttctcaattc accgttcaat tcgaagtaga cttctctttg
      361 accggttttc ttgttcaaat caccaacagc ttgtaatttg ataatcaaag tcttaccttg
      421 ttcaatggtg acttcgattt cttcgtctgg ttcagctgga gccaagaagt tcttggttgg
      481 taagacagac aagtcaccgt aagtttctct aatcttttgg aaatcttcgt agacacgagg
      541 atacatgttg taggaggcaa catcacattc atcgatatca ccgaatctgt tttgcaaatc
      601 ttctctgatc ttttccaaat cgaatggttc caattctaaa cctggtctac aagtcaattt
      661 acgacgcttg tttctcaaaa catcggatct caatggttct gggaaaccac cgtatggttg
      721 accgatcaaa ccttcgaaga agtccataac ggagtctggg aagtccaaag agttagctaa
      781 tcttctgatg tcatcagaag tcaatttgtt agaaaccatg aattgagcca aatcaccaac
      841 gaccttagaa gttggggtga ccttaacaat gtcacccaac aagtagttag cttctctgta
      901 agctctcttg gtttcagccc attgttcacc tagacctaat tgttgagctt ggaacaacaa
      961 gttggtcaat tgaccacctg gaatttcgtg ttggtaaact tcagggtctg gacccttcaa
     1021 gtcagcttcg aaacaagagt ataataatct catttcagcc cagtaagcgt ccaattctct
     1081 aacgtgttcg acgttgatac cagtgtcaat gttaccttcc aaggaggcca acaaagcgtt
     1141 aatagatggt tgagaggtca aaccggacat ggagttaatg gcaacgtcaa caacatcagc
     1201 accggccaaa gcacaagcag tcatggaagc aacagcagta ccagcggagt catgagagtg
     1261 aacgtggatt ggcaagtctg ggtatctggt acgtaaagaa ccaatcaaca atttggcagc
     1321 agctggcttc atggtaccag ccatgtcctt gatacccaag atgtgagtac ccatttgaac
     1381 gatcttttcg acaacttcta agtagtagtc caagttgtat ttcttacctg gttgcaacat
     1441 gtcaccagag taacaaacgg tagcttcgac aacaccacca gccttcttga cagcattaac
     1501 accaaccttc aattgttcca agtcgttcaa agcgtcaaag actctgaaaa tgtcaacacc
     1561 attgtccttg gcttgcttaa cgaaatggtc aatggcgttg tctggcaaag aggagtaagc
     1621 aacaccgtta gcacctctca acaacatttg gaatggaatg tttgggacca aagatctcaa
     1681 ttttctcaat ctttcccatg ggtcctcatg caagaatctc atggcgacat cgaaagtagc
     1741 accaccccaa cattccaaag caaaggcacc agccaaagcg tgagcagtgg ttggagcaat
     1801 ggtggccaaa tcgtgagttc tgactctggt agctagcaaa gattggtgag catctctcca
     1861 ggtggtgtcc atcaacaaag taccgttgaa ttgtctgact tgcttggcaa attcggatgg
     1921 acccttttcc aacaagactt gtctccaacc ggatggtggg gcagatttgg taacgttgat
     1981 gacattacct tgagcatcgt gcaaatgtgg aacggaaggg ttggacttca actttggtaa
     2041 accgatttga cccttgatag aggaaccgtt gacggccaag tcagccaagt agtgcaacaa
     2101 tttttgagct ctgttttgag aggaaaccat ttggaacaat tgtggagtgt catcgatgaa
     2161 agtggtccag taggtacctt caatgaaaac tgggttggtc aacaaagtca acaagaatgg
     2221 gatgttggtc ttgacacctc tgattctgaa ttcaatcaaa gctctgatca tctttcttct
     2281 gacaatttcg taggtagaac cagaacagga acacttaacc aacatggagt cgtagtgtgg
     2341 agagatggta gcaccagcgt aggcgttacc accgtccaat ctgacaccgt taccaccagc
     2401 ggatctgtag acttccaaac gaccagtgtc tggttggaag ttcttggatg gatcttcagt
     2461 ggtgatacga cattggatag agaaacctct ggtggtgatt ttgtcttgta atagacctaa
     2521 ttgagtcaaa gtagcaccag cggcgatttg gatttgagcg gagacaatgt caataccagt
     2581 gatttcttca gtgatggtgt gttcgacttg aattcttggg ttgatttcaa tgaagtaatg
     2641 tctgttttgg ttgtcaacca agaattcagc agtaccagcg tttctgtaac cacaaacctt
     2701 agccaactta acggcatcgg tcaagatagc atctctaact tctcttggta aagtcttagc
     2761 tggagcaact tcgacaacct tttggtgacg tctttggacg gaacagtctc tttcgaacaa
     2821 gtggacaacg ttaccgtggt tgtcagctaa taattgaact tcaatgtgct ttggcttgtc
     2881 caagaatctt tcgacgaaac aagtaccgtt accgaaagca gttctagctt cagaagtggc
     2941 tctttggaaa gcatcagcga catcgtcacc ttctctgaca actctcatac cacgaccacc
     3001 accaccgaaa gcagccttga tgataactgg gtaaccgtat tcattgacga aatctagagc
     3061 ttcttgaacg gtttcgatag gacctggagt acctgggaca gttggaacgt tggcacgagc
     3121 agccaaatgt ctggcagaaa ccttgtcacc aacagattca atgacttcag ctggtggacc
     3181 aatccaggta ataccagcct taacaacctt gtcagcaaat tcagagtttt cagacaagaa
     3241 accgtaacct gggtggatga aatcgacctt gtgcttcttg gcaatttcga tgatttcgtc
     3301 catggccaag taagcaccga ctggggtgta ttgaccttct tcaccgataa cgtaggcttc
     3361 atcagccttc aatctgtgca tggataatct atcttcgtga gagtagatgg cgatagttct
     3421 catagacaat tcgtgagcag atctgaaaat tctgattggg atttcacctc tgttggcgac
     3481 caaaatcttg ttcttttcac ccaacaaaga aaagttgtct ctcaaaccag ccaagatctt
     3541 ggaagaggac attgttttat atttgttgta aaaagtagat aattacttcc ttgatgatct
     3601 gtaaaaaaga gaaaaagaaa gcatctaaga acttgaaaaa ctacgaatta gaaaagacca
     3661 aatatgtatt tcttgcattg accaatttat gcaagtttat atatatgtaa atgtaagttt
     3721 cacgaggttc tactaaacta aaccaccccc ttggttagaa gaaaagagtg tgtgagaaca
     3781 ggctgttgtt gtcacacgat tcggacaatt ctgtttgaaa gagagagagt aacagtacga
     3841 tcgaacgaac tttgctctgg agatcacagt gggcatcata gcatgtggta ctaaaccctt
     3901 tcccgccatt ccagaacctt cgattgcttg ttacaaaacc tgtgagccgt cgctaggacc
     3961 ttgttgtgtg acgaaattgg aagctgcaat caataggaag acaggaagtc gagcgtgtct
     4021 gggttttttc agttttgttc tttttgcaaa caaatcacga gcgacggtaa tttctttctc
     4081 gataagaggc cacgtgcttt atgagggtaa catcaattca agaaggaggg aaacacttcc
     4141 tttttctggc ccgcacgcgg aatattggcg gaacggacac acgtggatac aaacctggac
     4201 aacgttcccc acttcagaag ttcctatact ttctagagaa taggaacttc actatagagt
     4261 cgaataaggg cgacaccccc taattagccc gggcgaaagg cccagtcttt cgactgagcc
     4321 tttcgtttta tttgatgcct ggcagttccc tactctcgca tggggagtcc ccacactacc
     4381 atcggcgcta cggcgtttca cttctgagtt cggcatgggg tcaggtggga ccaccgcgct
     4441 actgccgcca ggcaaacaag gggtgttatg agccatattc aggtataaat gggctcgcga
     4501 taatgttcag aattggttaa ttggttgtaa cactgacccc tatttgttta tttttctaaa
     4561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4621 gaaaaaggaa gaatatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     4681 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     4741 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     4801 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     4861 ttatgccact tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     4921 ccactgcgat ccccggaaaa acagcgttcc aggtattaga agaatatcct gattcaggtg
     4981 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcactcgatt cctgtttgta
     5041 attgtccttt taacagcgat cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata
     5101 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     5161 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     5221 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     5281 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     5341 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     5401 tgaataaatt gcaatttcat ttgatgctcg atgagttttt ctaagcagag cattacgctg
     5461 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     5521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5701 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
     5761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6181 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6241 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6301 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
     6361 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     6421 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     6481 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     6541 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     6601 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     6661 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     6721 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     6781 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     6841 ataagatgtt atcagtattt attatgcatt tagaataaat tttgtgtcgc ccttattcga
     6901 ctcactatag aagttcctat tctctagaaa gtataggaac ttcacttcat tttaggtaca
     6961 acaagcacga ccgagcatat gggaaggatg tttgtggtta tttgaggtga gtaacccata
     7021 tagagatcgt acacatttta caaggattta gagacaaagt aatattcttc gttgatatag
     7081 aggtgttcaa ttgttaaatg cttttcttct ttttattaga aaaagcgcct tgctttttgt
     7141 tgtatcactt gtaaatctac cgtcccttac aagaacattc acgaaattta agtggctcag
     7201 aatgaaaaag aaacaatata aaaaagcttt ccgtagtcat caatttattt tacataacac
     7261 tagatataaa gaaaagaaga taatattttt atataatta
//
