LOCUS       sequence-254028-        4817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 1. ITE transcription unit with the HTA1 
            promoter and VMA2 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4817)
  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 4817)
  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-5a-Phta1-ITE-Tvma2
FEATURES             Location/Qualifiers
     source          1..4817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    117..150
                     /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(2241..2274)
                     /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(2481..2500)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2567..2658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2659..3468
                     /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      3590..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4079..4098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4332..4349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4447..4464
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4448..4464
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata agatgttatc
       61 agtatttatt atgcatttag aataaatttt gtgtcgccct tattcgactc actatagaag
      121 ttcctattct ctagaaagta taggaacttc acttcatttt aaagcaaagg aaggagagaa
      181 cagaggagta cttgtacgtt cgatgggcaa gaggtgtgtt ccttgatctt tttctttttt
      241 ttttttcaat tacggattat gcatatatag gtgggtatac gtacgtaaaa aaacaaaaac
      301 acacatgtag acatctatat acgtttaaaa aagctttaga ttcagatctt tgggttttac
      361 aaaaaaaaaa cggacaaaat aaaaaaagcc tttttcttca gcaaccgtcc ttttattgtg
      421 gacaggtaca caatgggaac aagtagatca aagtgtggaa taaaccagca acgtaggtgt
      481 tcaaacagtt gaaagaaccc atcaaacccc aaccagcaaa gaaagcagca atgattggaa
      541 caaccatgtc acccttgtcc tcttgcaaag tccaaccgta gatcaaagta ccagctggca
      601 agacagcaaa caaagtgatc aaaccagagt gcaatctgtc ttgaggtaat ctgaaacctc
      661 tcttgacaat gtatcttctg acggttctgt cagacaattt accaccaacc aaagaaccaa
      721 tcaagaaacc agcacctgga gccaagtaga ataaaccgga aaccaaagcg gtggtcaaat
      781 ggaatctgga gttgaagata gctctggcag aagtcaagat ggagtattga gtgatggcca
      841 acaaaccaca acacaaatca gccaaaaaga catttgggta aacccattgc ttcaagacat
      901 cggttggaga gaacttggag atgatggtga ccaaagtggt tggcttgaaa gcagtagaaa
      961 ccttttcaga gttaccttcg atctttggga agaataatag agacaaaacc aaacctaaac
     1021 cagacatacc caattgtaac cagaagataa ctctccagga ggtgaaagtg acaatgacac
     1081 caccgacaca tggaccaatg gctggaccgg atagagtacc agccatgaag aaaccgacag
     1141 cagtacctct gtaaactggt tcaaagatat cagccaagac ggtttgaccg gaaaccatga
     1201 aagaagtacc ggtcaagcca gacaagacac ggaaagcaat gaacatttct tcgttgatgg
     1261 cagcagcagt accagcagaa caggcacaca acatggagat ggccaagttg taagaggttc
     1321 ttctaccaac caatttgttc attggacccc agatcaaaga agagtaaccc atggcaacca
     1381 aaacaccagc gttggaaatg ttaatggttt cgacggtcat atcgaattcg ttagcaattt
     1441 ctggagcagc tggcaacaaa caggtggagg agaaagtgac aaccaaagtc atccaggaaa
     1501 caacgaaagt gataacacac tttctccata atggaatgtc acgacccttc ttttcaggtt
     1561 cgttttgacc agaaccttca gtggtggtgg ttgggtttgg agattcagtg tcaccgtgac
     1621 ccattttata ttttatatgt atgaaatttg tttgttttga agttgtttat tcactgagaa
     1681 ataaccaaat ccgtatgatg atgtagtatc aagaagagaa gtacagattg gaagtaaata
     1741 gatgatggtt caacaagacc agaaaatcta caagctgatt aggagactta tttatatatt
     1801 ttttaggtca agacttattg ctagtattta cgatccactg gctggcttcg tgaacgggga
     1861 agggggtgag aaaagatttt gaaatcaaca aagtgggcaa taacaaataa cagcatgaga
     1921 aaccacatat ctctacgggc gtttcttcaa caacgacgag ttaactattg tgctcttttt
     1981 ttgagccacc aaatacactc cattccaata gcttcgcaca gtgaggcgaa aattttggaa
     2041 cagcgctaat gaattatttg tgagctcggc gagttcaaat ttgaagaaaa cgcggttggg
     2101 tcgttaacta tggttagacg ctcaatgtcg cccgaaaggg aaggctgttc tcactttttc
     2161 gcgcgttgca ccctttgcac aaacgcctgt gggtgtggta ctggatatgc aaagcgattg
     2221 gaagtcgctt ccccacttca gaagttccta tactttctag agaataggaa cttcactata
     2281 gagtcgaata agggcgacac cccctaatta gcccgggcga aaggcccagt ctttcgactg
     2341 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gtccccacac
     2401 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
     2461 cgctactgcc gccaggcaaa caaggggtgt tatgagccat attcaggtat aaatgggctc
     2521 gcgataatgt tcagaattgg ttaattggtt gtaacactga cccctatttg tttatttttc
     2581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2641 tattgaaaaa ggaagaatat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     2701 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2761 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2821 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2881 gaatttatgc cacttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2941 ctcaccactg cgatccccgg aaaaacagcg ttccaggtat tagaagaata tcctgattca
     3001 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcactc gattcctgtt
     3061 tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg
     3121 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3181 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     3241 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3301 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3361 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3421 gatatgaata aattgcaatt tcatttgatg ctcgatgagt ttttctaagc agagcattac
     3481 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgcg
     3541 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgttctt ctagtgtagc cgtagttagc ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4201 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4261 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4321 cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg cgagagtagg gaactgccag
     4381 gcatcaaact aagcagaagg cccctgacgg atggcctttt tgcgtttcta caaactcttt
     4441 ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc ctgggcggtt ctgataacga
     4501 gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg cgggtttttt tatgggggga
     4561 gtttagggaa agagcatttg tcagaatatt taagggcgcc tgtcactttg cttgatatat
     4621 gagaattatt taaccttata aatgagaaaa aagcaacgca ctttaaataa gatacgttgc
     4681 tttttcgatt gatgaacacc tataattaaa ctattcatct attatttatg attttttgta
     4741 tatacaatat ttctagtttg ttaaagagaa ttaagaaaat aaatctcgaa aataataaag
     4801 ggaaaatcag tttttga
//
