LOCUS       sequence-254118-        7087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 5. PYC transcription unit with the PXR1 
            promoter and NAT1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7087)
  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 7087)
  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-Ppxr1-PYC-Tnat1
FEATURES             Location/Qualifiers
     source          1..7087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(3627..3660)
                     /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(4130..4147)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4130..4146)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4245..4262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4416..5004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4496..4515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5126..5935)
                     /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(5936..6027)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6094..6113
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6320..6353
                     /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 acgaattgtc tatgagaact atcgccatct actctcacga agatagatta tccatgcaca
       61 gattgaaggc tgatgaagcc tacgttatcg gtgaagaagg tcaatacacc ccagtcggtg
      121 cttacttggc catggacgaa atcatcgaaa ttgccaagaa gcacaaggtc gatttcatcc
      181 acccaggtta cggtttcttg tctgaaaact ctgaatttgc tgacaaggtt gttaaggctg
      241 gtattacctg gattggtcca ccagctgaag tcattgaatc tgttggtgac aaggtttctg
      301 ccagacattt ggctgctcgt gccaacgttc caactgtccc aggtactcca ggtcctatcg
      361 aaaccgttca agaagctcta gatttcgtca atgaatacgg ttacccagtt atcatcaagg
      421 ctgctttcgg tggtggtggt cgtggtatga gagttgtcag agaaggtgac gatgtcgctg
      481 atgctttcca aagagccact tctgaagcta gaactgcttt cggtaacggt acttgtttcg
      541 tcgaaagatt cttggacaag ccaaagcaca ttgaagttca attattagct gacaaccacg
      601 gtaacgttgt ccacttgttc gaaagagact gttccgtcca aagacgtcac caaaaggttg
      661 tcgaagttgc tccagctaag actttaccaa gagaagttag agatgctatc ttgaccgatg
      721 ccgttaagtt ggctaaggtt tgtggttaca gaaacgctgg tactgctgaa ttcttggttg
      781 acaaccaaaa cagacattac ttcattgaaa tcaacccaag aattcaagtc gaacacacca
      841 tcactgaaga aatcactggt attgacattg tctccgctca aatccaaatc gccgctggtg
      901 ctactttgac tcaattaggt ctattacaag acaaaatcac caccagaggt ttctctatcc
      961 aatgtcgtat caccactgaa gatccatcca agaacttcca accagacact ggtcgtttgg
     1021 aagtctacag atccgctggt ggtaacggtg tcagattgga cggtggtaac gcctacgctg
     1081 gtgctaccat ctctccacac tacgactcca tgttggttaa gtgttcctgt tctggttcta
     1141 cctacgaaat tgtcagaaga aagatgatca gagctttgat tgaattcaga atcagaggtg
     1201 tcaagaccaa catcccattc ttgttgactt tgttgaccaa cccagttttc attgaaggta
     1261 cctactggac cactttcatc gatgacactc cacaattgtt ccaaatggtt tcctctcaaa
     1321 acagagctca aaaattgttg cactacttgg ctgacttggc cgtcaacggt tcctctatca
     1381 agggtcaaat cggtttacca aagttgaagt ccaacccttc cgttccacat ttgcacgatg
     1441 ctcaaggtaa tgtcatcaac gttaccaaat ctgccccacc atccggttgg agacaagtct
     1501 tgttggaaaa gggtccatcc gaatttgcca agcaagtcag acaattcaac ggtactttgt
     1561 tgatggacac cacctggaga gatgctcacc aatctttgct agctaccaga gtcagaactc
     1621 acgatttggc caccattgct ccaaccactg ctcacgcttt ggctggtgcc tttgctttgg
     1681 aatgttgggg tggtgctact ttcgatgtcg ccatgagatt cttgcatgag gacccatggg
     1741 aaagattgag aaaattgaga tctttggtcc caaacattcc attccaaatg ttgttgagag
     1801 gtgctaacgg tgttgcttac tcctctttgc cagacaacgc cattgaccat ttcgttaagc
     1861 aagccaagga caatggtgtt gacattttca gagtctttga cgctttgaac gacttggaac
     1921 aattgaaggt tggtgttaat gctgtcaaga aggctggtgg tgttgtcgaa gctaccgttt
     1981 gttactctgg tgacatgttg caaccaggta agaaatacaa cttggactac tacttagaag
     2041 ttgtcgaaaa gatcgttcaa atgggtactc acatcttggg tatcaaggac atggctggta
     2101 ccatgaagcc agctgctgcc aaattgttga ttggttcttt acgtaccaga tacccagact
     2161 tgccaatcca cgttcactct catgactccg ctggtactgc tgttgcttcc atgactgctt
     2221 gtgctttggc cggtgctgat gttgttgacg ttgccattaa ctccatgtcc ggtttgacct
     2281 ctcaaccatc tattaacgct ttgttggcct ccttggaagg taacattgac actggtatca
     2341 acgtcgaaca cgttagagaa ttggacgctt actgggctga aatgagatta ttatactctt
     2401 gtttcgaagc tgacttgaag ggtccagacc ctgaagttta ccaacacgaa attccaggtg
     2461 gtcaattgac caacttgttg ttccaagctc aacaattagg tctaggtgaa caatgggctg
     2521 aaaccaagag agcttacaga gaagctaact acttgttggg tgacattgtt aaggtcaccc
     2581 caacttctaa ggtcgttggt gatttggctc aattcatggt ttctaacaaa ttgacttctg
     2641 atgacatcag aagattagct aactctttgg acttcccaga ctccgttatg gacttcttcg
     2701 aaggtttgat cggtcaacca tacggtggtt tcccagaacc attgagatcc gatgttttga
     2761 gaaacaagcg tcgtaaattg acttgtagac caggtttaga attggaacca ttcgatttgg
     2821 aaaagatcag agaagatttg caaaacagat tcggtgatat cgatgaatgt gatgttgcct
     2881 cctacaacat gtatcctcgt gtctacgaag atttccaaaa gattagagaa acttacggtg
     2941 acttgtctgt cttaccaacc aagaacttct tggctccagc tgaaccagac gaagaaatcg
     3001 aagtcaccat tgaacaaggt aagactttga ttatcaaatt acaagctgtt ggtgatttga
     3061 acaagaaaac cggtcaaaga gaagtctact tcgaattgaa cggtgaattg agaaagatca
     3121 gagttgctga caaatctcaa aacattcaat ctgttgccaa gccaaaggct gatgtccacg
     3181 acacccacca aatcggtgct ccaatggctg gtgtcatcat tgaagtcaag gttcacaagg
     3241 gttctttggt caagaagggt gaatctatcg ccgttttgtc tgctatgaag atggaaatgg
     3301 ttgtttcctc tccagctgat ggtcaagtca aagatgtctt tatccgtgac ggtgaatccg
     3361 tcgatgcttc tgacttgttg gttgttttgg aagaagaaac tctaccacct tctcaaaaga
     3421 aataaactgc aactcctcaa tgtgtcaatt aactcttact taatttatgt atatattttt
     3481 tatgtatatg cttatatgca tgcgcatatg ctcataaaag atacattgtt ataggtcatt
     3541 tcttttccaa gctacatcta gccctcaaat aaccacaaac atccttccca tatgctcggt
     3601 cgtgcttgtt gtacctaaaa tgaagtgaag ttcctatact ttctagagaa taggaacttc
     3661 tatagtgagt cgaataaggg cgacacaaaa tttattctaa atgcataata aatactgata
     3721 acatcttata gtttgtatta tattttgtat tatcgttgac atgtataatt ttgatatcaa
     3781 aaactgattt tccctttatt attttcgaga tttattttct taattctctt taacaaacta
     3841 gaaatattgt atatacaaaa aatcataaat aatagatgaa tagtttaatt ataggtgttc
     3901 atcaatcgaa aaagcaacgt atcttattta aagtgcgttg cttttttctc atttataagg
     3961 ttaaataatt ctcatatatc aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc
     4021 tctttcccta aactcccccc ataaaaaaac ccgccgaagc gggtttttac gttatttgcg
     4081 gattaacgat tactcgttat cagaaccgcc cagggggccc gagcttaaga ctggccgtcg
     4141 ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc catccgtcag gggccttctg
     4201 cttagtttga tgcctggcag ttccctactc tcgccttccg cttcctcgct cactgactcg
     4261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4801 gtaggcggtg ctacagagtt cttgaagtgg tgggctaact acggctacac tagaagaaca
     4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5041 cagtggaacg acgcgcgcgt aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc
     5101 ccgtcaagtc agcgtaatgc tctgcttaga aaaactcatc gagcatcaaa tgaaattgca
     5161 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     5221 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     5281 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     5341 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     5401 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     5461 ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa
     5521 aaggacaatt acaaacagga atcgagtgca accggcgcag gaacactgcc agcgcatcaa
     5581 caatattttc acctgaatca ggatattctt ctaatacctg gaacgctgtt tttccgggga
     5641 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     5701 gtggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     5761 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaagcgat
     5821 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     5881 catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg ctcatattct
     5941 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6001 ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc
     6061 tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc ttgtttgcct
     6121 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     6181 agcgccgatg gtagtgtggg gactccccat gcgagagtag ggaactgcca ggcatcaaat
     6241 aaaacgaaag gctcagtcga aagactgggc ctttcgcccg ggctaattag ggggtgtcgc
     6301 ccttattcga ctctatagtg aagttcctat tctctagaaa gtataggaac ttctgaagtg
     6361 gggaacgttg tccaggtttg tatccacgtg tgtccgttcc gccaatattc cgcgtgctaa
     6421 aaacgcagca actaaagaag gcgactaata aaaattgagt atgcctctcc ttagcgtaaa
     6481 cagtaactgc gtgctataag gtggcactaa acttccttct tcccctttct atcggatttg
     6541 tgcagccagg agggatacca gtgacggcta tagcgggaga aagacgggcc ctctcacaac
     6601 cctttgcccc ttgcggttag tcatatagat tttgcttgtg aagagaactg gtcggttcaa
     6661 agatgctttc ttcaaagtac tatagccttg cccactctct ttctcccttc tgatctcttg
     6721 tatgctgatt ttactttaat aagtgaaggt gatcaatcag tcaactaaca aattagccgc
     6781 cactgcaagg atattttctc tgtcgttgtt gttttcctac aaatataaac aaccgggtaa
     6841 cagtcaaaga aaaaaaaaaa aagaaaagga aaaaaaaaaa atatgaaaaa ttttgttctg
     6901 aaagaagcga tgagatgagt actcaatagt aacatatagg cagcttacac cattaaacaa
     6961 agaagcagaa tgtcctcttc caagatcttg gctggtttga gagacaactt ttctttgttg
     7021 ggtgaaaaga acaagatttt ggtcgccaac agaggtgaaa tcccaatcag aattttcaga
     7081 tctgctc
//
