LOCUS       sequence-254056-        5004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 3. CAD transcription unit with the EFT2 
            promoter and K. lactis URA3 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5004)
  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 5004)
  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-bc-Peft2-CAD-Tklura3
FEATURES             Location/Qualifiers
     source          1..5004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    90..123
                     /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(2401..2434)
                     /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(2904..2921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2904..2920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3019..3036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3190..3778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3270..3289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3900..4709)
                     /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(4710..4801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4868..4887
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgcccg ggctaattag
       61 ggggtgtcgc ccttattcga ctctatagtg aagttcctat tctctagaaa gtataggaac
      121 ttctgaagtg gggcggatcg atgtacacaa ccgactgcac ccaaacgaac acaaatctta
      181 gcagtgcttc cttttttttc tttttttttt accagaagaa ttacgtacaa aagtacctac
      241 tatttcaaag caagaaatga gatgcctatt gtggttatat acagaataga tgataaatgg
      301 gttttccgtg caaaacgata tggagaattc aaaatgggtg cgaaatacct ggaacgtaag
      361 cgttctgaga aatacacaga cgcattaacc tgacaaaaac acaactagtt tgggaaaggg
      421 atttggtctt tcctctcggg cttctcgtgt ggttcctttc tttctcagat ctccctgcac
      481 actgggctgt tgtcctccag gttatggttt gttctcttca ggtattacaa tgcagtaggc
      541 ttttggagtg agcaaaacga agagagaaaa aaattttttc ttaaaagttt tttttcattt
      601 tgtgagctta ttcttctttt ctatatattc ttgatatctt agattataca tattattctc
      661 ttacatttca cgattgccct tttggtgttt agcattcagt ctcaaagacc acaaacacaa
      721 actataacat aattgcaaaa tgaccaagca atctgctgac tccaatgcca agtctggtgt
      781 cacttccgaa atctgtcact gggcttccaa cttggctact gacgacattc catctgatgt
      841 cttggaaaga gccaagtact tgattttgga cggtattgcc tgtgcttggg ttggtgctcg
      901 tgttccatgg tctgaaaagt acgttcaagc taccatgtcc ttcgaacctc caggtgcttg
      961 tcgtgtcatc ggttacggtc aaaaattggg tccagttgct gctgccatga ccaactctgc
     1021 ctttattcaa gccactgaat tggatgacta ccactctgaa gctccattgc actctgcttc
     1081 cattgttcta ccagctgttt tcgctgcttc tgaagtcttg gctgaacaag gtaagaccat
     1141 ctctggtatc gatgttatct tagctgccat tgtcggtttc gaatctggtc caagaatcgg
     1201 taaggccatc tacggttctg acttgttgaa caacggttgg cattgtggtg ccgtttacgg
     1261 tgctccagct ggtgctttgg ctaccggtaa gttgttgggt ttgactccag actccatgga
     1321 agatgctttg ggtatcgctt gtacccaagc ttgtggtttg atgtctgctc aatacggtgg
     1381 tatggttaag agagttcaac atggtttcgc tgccagaaac ggtctattag gtggtttgtt
     1441 ggctcacggt ggttacgaag ctatgaaggg tgtcttggaa agatcttacg gtggtttctt
     1501 gaagatgttc accaagggta acggtagaga acctccatac aaggaagaag aagttgttgc
     1561 cggtttaggt tctttctggc acactttcac catcagaatc aaattgtacg cttgttgtgg
     1621 tttagtccac ggtccagttg aagccattga aaacttacaa ggtcgttacc cagaattgtt
     1681 gaacagagct aacttgtcca acatcagaca cgttcacgtt caattatcca ctgcttccaa
     1741 ctctcactgt ggttggattc cagaagaaag accaatctcc tccattgctg gtcaaatgtc
     1801 tgttgcttac attttggctg tccaattggt tgaccaacaa tgtttgttgt ctcaattctc
     1861 cgaattcgat gacaacttgg aaagaccaga agtctgggat ttggctagaa aggtcacctc
     1921 ttctcaatct gaagaatttg accaagatgg taactgtttg tctgctggta gagtcagaat
     1981 tgaattcaac gacggttctt ccatcactga atccgttgaa aagccattag gtgtcaagga
     2041 accaatgcca aacgaaagaa tcttgcacaa atacagaact ttggctggtt ccgtcactga
     2101 cgaatccaga gtcaaggaaa tcgaagattt ggttttgggt ttggacagat tgaccgatat
     2161 ctctccatta ttggaattgt tgaactgtcc agtcaaatct ccattgggta tcaagtaaat
     2221 atacaggaaa cttaatagaa caaatcacat atttaatcta atagccacct gcattggcac
     2281 ggtgcaacac tacttcaact tcatcttaca aaaagatcac gtgatctgtt gtattgcctc
     2341 acgctttccg gcatcttcca gaccacagta tatccatccg cctcctgttg aaaatgaagt
     2401 gaagttccta tactttctag agaataggaa cttctatagt gagtcgaata agggcgacac
     2461 aaaatttatt ctaaatgcat aataaatact gataacatct tatagtttgt attatatttt
     2521 gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt tattattttc
     2581 gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac aaaaaatcat
     2641 aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca acgtatctta
     2701 tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata tatcaagcaa
     2761 agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc ccccataaaa
     2821 aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg ttatcagaac
     2881 cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa gagtttgtag
     2941 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct
     3001 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3601 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca
     3841 cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgct
     3901 tagaaaaact catcgagcat caaatgaaat tgcaatttat tcatatcagg attatcaata
     3961 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     4021 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     4081 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     4141 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     4201 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     4261 gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgag
     4321 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     4381 tcttctaata cctggaacgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
     4441 tcaggagtac ggataaaatg cttgatggtc ggaagtggca taaattccgt cagccagttt
     4501 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     4561 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
     4621 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     4681 ctcgacgttt cccgttgaat atggctcata ttcttccttt ttcaatatta ttgaagcatt
     4741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4801 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
     4861 cctgaatatg gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc
     4921 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc
     4981 ccatgcgaga gtagggaact gcca
//
