LOCUS       sequence-254027-        4791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 1. ITE transcription unit with the ACT1 
            promoter and DPP1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  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 4791)
  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-Pact1-ITE-Tdpp1
FEATURES             Location/Qualifiers
     source          1..4791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    472..505
                     /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(2570..2603)
                     /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(2810..2829)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2896..2987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2988..3797
                     /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      3919..4507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4408..4427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4661..4678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     join(4776..4791,1..2)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(4777..4791,1..2)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
       61 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
      121 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
      181 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
      241 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
      301 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
      361 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
      421 ttattatgca tttagaataa attttgtgtc gcccttattc gactcactat agaagttcct
      481 attctctaga aagtatagga acttcacttc attttaaagc aaaggaagga gagaacagag
      541 gagtacttgt acgttcgatg ggcaagaggc gtgtaaagtg atgttgggat tgtccgatga
      601 aataacaagc aggcccttgc acgtcaagat taatttgaat aaagaagtaa catgtcatct
      661 ttatgtaatc gctgttattc catacagaac aataaatacg tatatttcgt atgtcatgtg
      721 gagtatatat tcttttttat ttttattgtg gacaggtaca caatgggaac aagtagatca
      781 aagtgtggaa taaaccagca acgtaggtgt tcaaacagtt gaaagaaccc atcaaacccc
      841 aaccagcaaa gaaagcagca atgattggaa caaccatgtc acccttgtcc tcttgcaaag
      901 tccaaccgta gatcaaagta ccagctggca agacagcaaa caaagtgatc aaaccagagt
      961 gcaatctgtc ttgaggtaat ctgaaacctc tcttgacaat gtatcttctg acggttctgt
     1021 cagacaattt accaccaacc aaagaaccaa tcaagaaacc agcacctgga gccaagtaga
     1081 ataaaccgga aaccaaagcg gtggtcaaat ggaatctgga gttgaagata gctctggcag
     1141 aagtcaagat ggagtattga gtgatggcca acaaaccaca acacaaatca gccaaaaaga
     1201 catttgggta aacccattgc ttcaagacat cggttggaga gaacttggag atgatggtga
     1261 ccaaagtggt tggcttgaaa gcagtagaaa ccttttcaga gttaccttcg atctttggga
     1321 agaataatag agacaaaacc aaacctaaac cagacatacc caattgtaac cagaagataa
     1381 ctctccagga ggtgaaagtg acaatgacac caccgacaca tggaccaatg gctggaccgg
     1441 atagagtacc agccatgaag aaaccgacag cagtacctct gtaaactggt tcaaagatat
     1501 cagccaagac ggtttgaccg gaaaccatga aagaagtacc ggtcaagcca gacaagacac
     1561 ggaaagcaat gaacatttct tcgttgatgg cagcagcagt accagcagaa caggcacaca
     1621 acatggagat ggccaagttg taagaggttc ttctaccaac caatttgttc attggacccc
     1681 agatcaaaga agagtaaccc atggcaacca aaacaccagc gttggaaatg ttaatggttt
     1741 cgacggtcat atcgaattcg ttagcaattt ctggagcagc tggcaacaaa caggtggagg
     1801 agaaagtgac aaccaaagtc atccaggaaa caacgaaagt gataacacac tttctccata
     1861 atggaatgtc acgacccttc ttttcaggtt cgttttgacc agaaccttca gtggtggtgg
     1921 ttgggtttgg agattcagtg tcaccgtgac ccattgttaa ttcagtaaat tttcgatctt
     1981 gggaagaaaa aagcagtaag cgtgaaaaat ctaaaagctg atgtagtaga agatcctatt
     2041 ctttaacaaa gattgacctt ttctttttct tcttggtttg agtagaaagg ggaaggaaga
     2101 atacaagaga gaggaaaaaa aggaagataa aaagagagcg tgatataaat gaatatatat
     2161 taaacaagag agattgggaa ggaaaggatc aaacaaaccc aaaaatattt caaaaaggag
     2221 agagagaggc gagtttggtt tcaaaacggt ttatttattt atgcaagagg acgtggaaga
     2281 aaaagaagaa agaagaaaaa aatttgaaag aaaaaaacgc gtggcgggta aagaagaaaa
     2341 tggaaaatag aggccgggtg acagagaaat attgagggtt aattggaaaa tatgttaggg
     2401 tgaggcatat gtttttaagg gttttgagga tccgataagg aagaatgtag gttaaatgtt
     2461 gtgcattaat tgctgtggca gcttacccgc ttccccacac attttgcaca aacgcctgtg
     2521 ggtgtggtac tggatatgca aagcgattgg aagtcgcttc cccacttcag aagttcctat
     2581 actttctaga gaataggaac ttcactatag agtcgaataa gggcgacacc ccctaattag
     2641 cccgggcgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggcagtt
     2701 ccctactctc gcatggggag tccccacact accatcggcg ctacggcgtt tcacttctga
     2761 gttcggcatg gggtcaggtg ggaccaccgc gctactgccg ccaggcaaac aaggggtgtt
     2821 atgagccata ttcaggtata aatgggctcg cgataatgtt cagaattggt taattggttg
     2881 taacactgac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2941 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatatg agccatattc
     3001 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3061 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3121 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3181 atgagatggt cagactaaac tggctgacgg aatttatgcc acttccgacc atcaagcatt
     3241 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcgt
     3301 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3361 tcctgcgccg gttgcactcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3421 ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3481 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3541 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3601 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3661 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3721 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcaattt catttgatgc
     3781 tcgatgagtt tttctaagca gagcattacg ctgacttgac gggacggcgc aagctcatga
     3841 ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga
     3901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4081 gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4681 agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc ccctgacgga
     4741 tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc a
//
