LOCUS       sequence-254127-        4534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 6. CSC transcription unit with the PFY1 
            promoter and AIP1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4534)
  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 4534)
  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-ef-Ppfy1-CSC-Taip1
FEATURES             Location/Qualifiers
     source          1..4534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    112..145
                     /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(1953..1986)
                     /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(2193..2212)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2279..2370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2371..3180
                     /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      3302..3890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3791..3810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4044..4061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4159..4176
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4160..4176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
       61 ttattatgca tttagaataa attttgtgtc gcccttattc gactcactat agaagttcct
      121 attctctaga aagtatagga acttcacttc attttgtgca ctacatcttg tgggtgtcga
      181 catgctggct ggattcgaag gtttcgagga ttcgggtaat aattacaagt ttatatggac
      241 tgtggtaaaa acagtgtaaa acttttttat atctgttttt ctaaggttac gtaattgttt
      301 actttatatt cttattttat ctcttgatat cagatttgaa atctctcttt tcgtaaccag
      361 tgtataattg tcttggtctg gcaatcttca taccatcaga gtgcatttcg gaccagtgag
      421 cgatccaacc aacagttctg gccatagcaa agataacggt gaacatggaa gatggaatac
      481 ccatagcctt taagatgata ccggagtaga aatcgacgtt tgggtacaat ttcttttcga
      541 tgaagtatgg gtcgttcaaa gcaatgtttt ccaattccat ggcaacttcc aacaagtcat
      601 ccttggtacc caattccttc aaaacttcgt gacaggtttc tctcatgacg gtagctcttg
      661 ggtcgtagtt cttgtagaca cggtgaccga aacccatcaa tctgaaagag tcgttcttgt
      721 ccttagctct tctgacaaat tctggaatgt gcttgacaga agaaatttct tccaacatct
      781 tcaaagcagc ttcgttggca ccaccatgag ctggacccca taaagaagca ataccagcag
      841 cgatacaagc aaatgggtta gcaccagaag aaccggcagt tctaacagta gaagtagaag
      901 cgttttgttc atggtcagcg tgcaagatca agattctgtc catggctctt tccaaaattg
      961 ggttaacttc gtaaggttca catggagtgg agaacatcat gttcaagaag ttaccggcgt
     1021 aagacaaatc gtttcttggg taaacgaaag gttgaccgat ggagtacttg taacacatag
     1081 cagccatggt tggcatcttg gacaacaaca agaaagcggc aatttctctg tgtcttgggt
     1141 tgttgacatc caaagagtca tggtagaaag cagccaaagc accagtgata ccacacatga
     1201 cagccattgg gtgggagtca cgacggaaag cgtggaataa tctggtgatt tgttcgtgaa
     1261 tcatggtgtg tctggtaacg gtggttttaa attcgtcgta ttgttcttga gttggctttt
     1321 caccattcaa caagatgtaa caaacttcca agtagttgga gtcagtagct aattggtcaa
     1381 ttgggaaacc tctgtgtaat aggatacctt cgtcaccatc gatgaaagtg atcttggatt
     1441 cacaagaagc agtagaggtg aaacctgggt cgaaggtgaa aacacccttg gaacccaaag
     1501 ttctgatatc aatgacatct tgacccaaag tacctttcaa aacatccaat tcgacagcag
     1561 tgtcaccgtt caaggtcaac ttggccttgg tgtcagccat tatttgcgat cgtagttggg
     1621 tttatgttgt gtatgtaatt tatcttttct caaggatcac acagttgttt taagaaggga
     1681 atgagaaagg gtcaactgag ctttaaaaag aaaaaaaata ttaatgcggg taatgagttt
     1741 cgtacaaaga catttcatgg cgtgttctcg ccaaaaatca tggtaaaacc tcccacacag
     1801 taacctcata ctaaatacta caacatgtca tcactccatt tctttgcgat tgtacatact
     1861 aatatatata aacaaagtaa cgtctcctgc acaggtacaa caagcacgac cgagcatatg
     1921 ggaaggatgt ttgtggttat ttccccactt cagaagttcc tatactttct agagaatagg
     1981 aacttcacta tagagtcgaa taagggcgac accccctaat tagcccgggc gaaaggccca
     2041 gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcatggg
     2101 gagtccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc atggggtcag
     2161 gtgggaccac cgcgctactg ccgccaggca aacaaggggt gttatgagcc atattcaggt
     2221 ataaatgggc tcgcgataat gttcagaatt ggttaattgg ttgtaacact gacccctatt
     2281 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2341 atgcttcaat aatattgaaa aaggaagaat atgagccata ttcaacggga aacgtcgagg
     2401 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2461 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2521 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2581 aactggctga cggaatttat gccacttccg accatcaagc attttatccg tactcctgat
     2641 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cgttccaggt attagaagaa
     2701 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcac
     2761 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgcct cgctcaggcg
     2821 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2881 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2941 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3001 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     3061 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     3121 attgataatc ctgatatgaa taaattgcaa tttcatttga tgctcgatga gtttttctaa
     3181 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3241 gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3421 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta gcccaccact
     3481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3661 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3721 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3781 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3841 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3901 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3961 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4021 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa ggcgagagta
     4081 gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac ggatggcctt tttgcgtttc
     4141 tacaaactct ttctgtgttg taaaacgacg gccagtctta agctcgggcc ccctgggcgg
     4201 ttctgataac gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt
     4261 tttatggggg gagtttaggg aaagagcatt tgtcagaata tttaagggcg cctgtcactt
     4321 tgcttgatat atgagaatta tttaacctta taaatgagaa aaaagcaacg cactttaaat
     4381 aagatacgtt gctttttcga ttgatgaaca cctataatta aactattcat ctattattta
     4441 tgattttttg tatatacaat atttctagtt tgttaaagag aattaagaaa ataaatctcg
     4501 aaaataataa agggaaaatc agtttttgat atca
//
