LOCUS       sequence-254124-        4934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 6. CSC transcription unit with the ZUO1 
            promoter and VMA16 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4934)
  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 4934)
  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-Pzuo1-CSC-Tvma16
FEATURES             Location/Qualifiers
     source          1..4934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..37
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     135..152
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     136..152
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    622..655
                     /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(2863..2896)
                     /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(3103..3122)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3189..3280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3281..4090
                     /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      4212..4800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4701..4720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga
       61 actgccaggc atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca
      121 aactctttct gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct
      181 gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta
      241 tggggggagt ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct
      301 tgatatatga gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga
      361 tacgttgctt tttcgattga tgaacaccta taattaaact attcatctat tatttatgat
      421 tttttgtata tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa
      481 taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat
      541 ataatacaaa ctataagatg ttatcagtat ttattatgca tttagaataa attttgtgtc
      601 gcccttattc gactcactat agaagttcct attctctaga aagtatagga acttcacttc
      661 attttgtgca ctacatcttg tgggtgtcga catgctggct ggattcgaag gtttcgagga
      721 ctgaatagct tttgggcaat atcaaccaga ataggaaaaa ttcaaaacca ttttaaataa
      781 atatatattt atatagttgt acattttttt ctcttactta atataatatg atttttagat
      841 aataaaagaa aacgtgcata tatacataca catgtatctc agatatctca ttatatctat
      901 tagtaaatat ggatgcgctt atctagctcg taaaaacgga aaagaaaagc ctggtttgag
      961 cgtttatctc ttgatatcag atttgaaatc tctcttttcg taaccagtgt ataattgtct
     1021 tggtctggca atcttcatac catcagagtg catttcggac cagtgagcga tccaaccaac
     1081 agttctggcc atagcaaaga taacggtgaa catggaagat ggaataccca tagcctttaa
     1141 gatgataccg gagtagaaat cgacgtttgg gtacaatttc ttttcgatga agtatgggtc
     1201 gttcaaagca atgttttcca attccatggc aacttccaac aagtcatcct tggtacccaa
     1261 ttccttcaaa acttcgtgac aggtttctct catgacggta gctcttgggt cgtagttctt
     1321 gtagacacgg tgaccgaaac ccatcaatct gaaagagtcg ttcttgtcct tagctcttct
     1381 gacaaattct ggaatgtgct tgacagaaga aatttcttcc aacatcttca aagcagcttc
     1441 gttggcacca ccatgagctg gaccccataa agaagcaata ccagcagcga tacaagcaaa
     1501 tgggttagca ccagaagaac cggcagttct aacagtagaa gtagaagcgt tttgttcatg
     1561 gtcagcgtgc aagatcaaga ttctgtccat ggctctttcc aaaattgggt taacttcgta
     1621 aggttcacat ggagtggaga acatcatgtt caagaagtta ccggcgtaag acaaatcgtt
     1681 tcttgggtaa acgaaaggtt gaccgatgga gtacttgtaa cacatagcag ccatggttgg
     1741 catcttggac aacaacaaga aagcggcaat ttctctgtgt cttgggttgt tgacatccaa
     1801 agagtcatgg tagaaagcag ccaaagcacc agtgatacca cacatgacag ccattgggtg
     1861 ggagtcacga cggaaagcgt ggaataatct ggtgatttgt tcgtgaatca tggtgtgtct
     1921 ggtaacggtg gttttaaatt cgtcgtattg ttcttgagtt ggcttttcac cattcaacaa
     1981 gatgtaacaa acttccaagt agttggagtc agtagctaat tggtcaattg ggaaacctct
     2041 gtgtaatagg ataccttcgt caccatcgat gaaagtgatc ttggattcac aagaagcagt
     2101 agaggtgaaa cctgggtcga aggtgaaaac acccttggaa cccaaagttc tgatatcaat
     2161 gacatcttga cccaaagtac ctttcaaaac atccaattcg acagcagtgt caccgttcaa
     2221 ggtcaacttg gccttggtgt cagccattgt tactcttgtt ggtatatgtg agacaaaacg
     2281 aagattttat aaaagcatcg gtaggaaaac cttatgaaat tttctctacc agctcttgaa
     2341 ataaaaattt cgaactcttt gcaaaatttc atcgagaaaa aaaaaagatt cgtatttgcg
     2401 ggtaatattt tttggacatt tgtattagga gcgttatgtt cattaatgtc atccaaaaga
     2461 tagttataaa tctatgattg tatggtttct ttgacatatt tatagttaat atatatatat
     2521 atatgtgttt aattttataa aaattccatc gtttatattc gattattttt atatgtcaac
     2581 ctttttttta agaaatgttc atatttagtc aagttcttaa tgaaatgaac ggttgaaagt
     2641 acaaggcaac tatcatttca agatataaaa aaattgccca aaaaaaaaag gccatttgat
     2701 gaaatgtctc atcgcgctac gatttctgct gtaatcagaa ctaaatgcgt gatcggaagt
     2761 aatattagat attggatgca taaaagataa aacgccaggc acaggtacaa caagcacgac
     2821 cgagcatatg ggaaggatgt ttgtggttat ttccccactt cagaagttcc tatactttct
     2881 agagaatagg aacttcacta tagagtcgaa taagggcgac accccctaat tagcccgggc
     2941 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact
     3001 ctcgcatggg gagtccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc
     3061 atggggtcag gtgggaccac cgcgctactg ccgccaggca aacaaggggt gttatgagcc
     3121 atattcaggt ataaatgggc tcgcgataat gttcagaatt ggttaattgg ttgtaacact
     3181 gacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3241 accctgataa atgcttcaat aatattgaaa aaggaagaat atgagccata ttcaacggga
     3301 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3361 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3421 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3481 ggtcagacta aactggctga cggaatttat gccacttccg accatcaagc attttatccg
     3541 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cgttccaggt
     3601 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3661 ccggttgcac tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgcct
     3721 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3781 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3841 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3901 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3961 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4021 aaaatatggt attgataatc ctgatatgaa taaattgcaa tttcatttga tgctcgatga
     4081 gtttttctaa gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     4141 cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     4201 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4261 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4321 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4381 gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4441 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4501 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4561 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4621 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4681 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4741 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4801 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4861 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4921 gataccgctc gccg
//
