LOCUS       sequence-254119-        7099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 5. PYC transcription unit with the FBA1 
            promoter and RPL15A terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7099)
  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 7099)
  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-de-Pfba1-PYC-Trpl15a
FEATURES             Location/Qualifiers
     source          1..7099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(3765..3798)
                     /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(4268..4285)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4268..4284)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4383..4400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4554..5142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4634..4653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5264..6073)
                     /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(6074..6165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6232..6251
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6458..6491
                     /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 attcaaaatg tcctcttcca agatcttggc tggtttgaga gacaactttt ctttgttggg
       61 tgaaaagaac aagattttgg tcgccaacag aggtgaaatc ccaatcagaa ttttcagatc
      121 tgctcacgaa ttgtctatga gaactatcgc catctactct cacgaagata gattatccat
      181 gcacagattg aaggctgatg aagcctacgt tatcggtgaa gaaggtcaat acaccccagt
      241 cggtgcttac ttggccatgg acgaaatcat cgaaattgcc aagaagcaca aggtcgattt
      301 catccaccca ggttacggtt tcttgtctga aaactctgaa tttgctgaca aggttgttaa
      361 ggctggtatt acctggattg gtccaccagc tgaagtcatt gaatctgttg gtgacaaggt
      421 ttctgccaga catttggctg ctcgtgccaa cgttccaact gtcccaggta ctccaggtcc
      481 tatcgaaacc gttcaagaag ctctagattt cgtcaatgaa tacggttacc cagttatcat
      541 caaggctgct ttcggtggtg gtggtcgtgg tatgagagtt gtcagagaag gtgacgatgt
      601 cgctgatgct ttccaaagag ccacttctga agctagaact gctttcggta acggtacttg
      661 tttcgtcgaa agattcttgg acaagccaaa gcacattgaa gttcaattat tagctgacaa
      721 ccacggtaac gttgtccact tgttcgaaag agactgttcc gtccaaagac gtcaccaaaa
      781 ggttgtcgaa gttgctccag ctaagacttt accaagagaa gttagagatg ctatcttgac
      841 cgatgccgtt aagttggcta aggtttgtgg ttacagaaac gctggtactg ctgaattctt
      901 ggttgacaac caaaacagac attacttcat tgaaatcaac ccaagaattc aagtcgaaca
      961 caccatcact gaagaaatca ctggtattga cattgtctcc gctcaaatcc aaatcgccgc
     1021 tggtgctact ttgactcaat taggtctatt acaagacaaa atcaccacca gaggtttctc
     1081 tatccaatgt cgtatcacca ctgaagatcc atccaagaac ttccaaccag acactggtcg
     1141 tttggaagtc tacagatccg ctggtggtaa cggtgtcaga ttggacggtg gtaacgccta
     1201 cgctggtgct accatctctc cacactacga ctccatgttg gttaagtgtt cctgttctgg
     1261 ttctacctac gaaattgtca gaagaaagat gatcagagct ttgattgaat tcagaatcag
     1321 aggtgtcaag accaacatcc cattcttgtt gactttgttg accaacccag ttttcattga
     1381 aggtacctac tggaccactt tcatcgatga cactccacaa ttgttccaaa tggtttcctc
     1441 tcaaaacaga gctcaaaaat tgttgcacta cttggctgac ttggccgtca acggttcctc
     1501 tatcaagggt caaatcggtt taccaaagtt gaagtccaac ccttccgttc cacatttgca
     1561 cgatgctcaa ggtaatgtca tcaacgttac caaatctgcc ccaccatccg gttggagaca
     1621 agtcttgttg gaaaagggtc catccgaatt tgccaagcaa gtcagacaat tcaacggtac
     1681 tttgttgatg gacaccacct ggagagatgc tcaccaatct ttgctagcta ccagagtcag
     1741 aactcacgat ttggccacca ttgctccaac cactgctcac gctttggctg gtgcctttgc
     1801 tttggaatgt tggggtggtg ctactttcga tgtcgccatg agattcttgc atgaggaccc
     1861 atgggaaaga ttgagaaaat tgagatcttt ggtcccaaac attccattcc aaatgttgtt
     1921 gagaggtgct aacggtgttg cttactcctc tttgccagac aacgccattg accatttcgt
     1981 taagcaagcc aaggacaatg gtgttgacat tttcagagtc tttgacgctt tgaacgactt
     2041 ggaacaattg aaggttggtg ttaatgctgt caagaaggct ggtggtgttg tcgaagctac
     2101 cgtttgttac tctggtgaca tgttgcaacc aggtaagaaa tacaacttgg actactactt
     2161 agaagttgtc gaaaagatcg ttcaaatggg tactcacatc ttgggtatca aggacatggc
     2221 tggtaccatg aagccagctg ctgccaaatt gttgattggt tctttacgta ccagataccc
     2281 agacttgcca atccacgttc actctcatga ctccgctggt actgctgttg cttccatgac
     2341 tgcttgtgct ttggccggtg ctgatgttgt tgacgttgcc attaactcca tgtccggttt
     2401 gacctctcaa ccatctatta acgctttgtt ggcctccttg gaaggtaaca ttgacactgg
     2461 tatcaacgtc gaacacgtta gagaattgga cgcttactgg gctgaaatga gattattata
     2521 ctcttgtttc gaagctgact tgaagggtcc agaccctgaa gtttaccaac acgaaattcc
     2581 aggtggtcaa ttgaccaact tgttgttcca agctcaacaa ttaggtctag gtgaacaatg
     2641 ggctgaaacc aagagagctt acagagaagc taactacttg ttgggtgaca ttgttaaggt
     2701 caccccaact tctaaggtcg ttggtgattt ggctcaattc atggtttcta acaaattgac
     2761 ttctgatgac atcagaagat tagctaactc tttggacttc ccagactccg ttatggactt
     2821 cttcgaaggt ttgatcggtc aaccatacgg tggtttccca gaaccattga gatccgatgt
     2881 tttgagaaac aagcgtcgta aattgacttg tagaccaggt ttagaattgg aaccattcga
     2941 tttggaaaag atcagagaag atttgcaaaa cagattcggt gatatcgatg aatgtgatgt
     3001 tgcctcctac aacatgtatc ctcgtgtcta cgaagatttc caaaagatta gagaaactta
     3061 cggtgacttg tctgtcttac caaccaagaa cttcttggct ccagctgaac cagacgaaga
     3121 aatcgaagtc accattgaac aaggtaagac tttgattatc aaattacaag ctgttggtga
     3181 tttgaacaag aaaaccggtc aaagagaagt ctacttcgaa ttgaacggtg aattgagaaa
     3241 gatcagagtt gctgacaaat ctcaaaacat tcaatctgtt gccaagccaa aggctgatgt
     3301 ccacgacacc caccaaatcg gtgctccaat ggctggtgtc atcattgaag tcaaggttca
     3361 caagggttct ttggtcaaga agggtgaatc tatcgccgtt ttgtctgcta tgaagatgga
     3421 aatggttgtt tcctctccag ctgatggtca agtcaaagat gtctttatcc gtgacggtga
     3481 atccgtcgat gcttctgact tgttggttgt tttggaagaa gaaactctac caccttctca
     3541 aaagaaataa actggttgat ggaaaatata attttattgg gcaaactttt gtttatctga
     3601 tgtgttttat actattatct ttttaattaa tgattctata tacaaacctg tatatttttt
     3661 ctttaaccaa tttttttttt tatagaccta gagctgtact cctcaaataa ccacaaacat
     3721 ccttcccata tgctcggtcg tgcttgttgt acctaaaatg aagtgaagtt cctatacttt
     3781 ctagagaata ggaacttcta tagtgagtcg aataagggcg acacaaaatt tattctaaat
     3841 gcataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     3901 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     3961 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     4021 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     4081 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     4141 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     4201 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     4261 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     4321 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     4381 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4441 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4501 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4561 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4621 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4681 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4741 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4801 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4861 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4921 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     4981 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5041 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5101 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5161 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     5221 tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc tgcttagaaa aactcatcga
     5281 gcatcaaatg aaattgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     5341 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     5401 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     5461 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     5521 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     5581 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgaggcgaa
     5641 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgagtgcaac cggcgcagga
     5701 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     5761 acgctgtttt tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     5821 aatgcttgat ggtcggaagt ggcataaatt ccgtcagcca gtttagtctg accatctcat
     5881 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     5941 gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     6001 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt
     6061 gaatatggct catattcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gtcagtgtta
     6181 caaccaatta accaattctg aacattatcg cgagcccatt tatacctgaa tatggctcat
     6241 aacacccctt gtttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     6301 tcagaagtga aacgccgtag cgccgatggt agtgtgggga ctccccatgc gagagtaggg
     6361 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgcccggg
     6421 ctaattaggg ggtgtcgccc ttattcgact ctatagtgaa gttcctattc tctagaaagt
     6481 ataggaactt ctgaagtggg gaacgttgtc caggtttgta tccacgtgtg tccgttccgc
     6541 caatattccg cgtgcatgac agcaggatta tcgtaatacg taatagttga aaatctcaaa
     6601 aatgtgtggg tcattacgta aataatgata ggaatgggat tcttctattt ttcctttttc
     6661 cattctagca gccgtcggga aaacgtggca tcctctcttt cgggctcaat tggtgtcacg
     6721 ctgccgtgag catcctctct ttccatatct aacaactgag cacgtaacca atggaaaagc
     6781 atgagcttag cgttgctcca aaaaagtatt ggatggttaa taccatttgt ctgttctctt
     6841 ctgactttgt ctcctcaaaa aaaaaaaatc tacaatcaac agatcgcttc aattacgccc
     6901 tcacaaaaac ttttttcctt cttcttcgcc cacgttaaat tttatccctc atgttgtcta
     6961 acggatttct gcacttgatt tattataaaa agacaaagac ataatacttc tctatcaatt
     7021 tcagttattg ttcttccttg cgttattctt ctgttcttct ttttcttttg tcatatataa
     7081 ccataaccaa gtaatacat
//
