LOCUS       sequence-254058-        4987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 3. CAD transcription unit with the S. mikatae
            TEF1 promoter and YHI9 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4987)
  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 4987)
  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-Psmtef1-CAD-Tyhi9
FEATURES             Location/Qualifiers
     source          1..4987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    45..78
                     /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(2339..2372)
                     /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(2579..2598)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2665..2756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2757..3566
                     /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      3688..4276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4177..4196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4430..4447
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4545..4562
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4546..4562
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcatttagaa taaattttgt gtcgccctta ttcgactcac tatagaagtt cctattctct
       61 agaaagtata ggaacttcac ttcattttca acaggaggcg gatggatata ctgtggtctg
      121 gaagatgccg gaaagcgtga gggagtaatt tggcttattg gtatgttgca aaaatactct
      181 ctgaaagaat acttcgaatt tcggcacttc ttatgtacac tttgatggaa gctttcttga
      241 taaggaattc tccgtctatt tccttcaata ccgcctccgg cggtattgaa ggagaaaaat
      301 ttgggcggga gcatattatt gtaattaata tatacatata tatatatata tatacatcaa
      361 ccttacaact atgcgtttag aattttactt gatacccaat ggagatttga ctggacagtt
      421 caacaattcc aataatggag agatatcggt caatctgtcc aaacccaaaa ccaaatcttc
      481 gatttccttg actctggatt cgtcagtgac ggaaccagcc aaagttctgt atttgtgcaa
      541 gattctttcg tttggcattg gttccttgac acctaatggc ttttcaacgg attcagtgat
      601 ggaagaaccg tcgttgaatt caattctgac tctaccagca gacaaacagt taccatcttg
      661 gtcaaattct tcagattgag aagaggtgac ctttctagcc aaatcccaga cttctggtct
      721 ttccaagttg tcatcgaatt cggagaattg agacaacaaa cattgttggt caaccaattg
      781 gacagccaaa atgtaagcaa cagacatttg accagcaatg gaggagattg gtctttcttc
      841 tggaatccaa ccacagtgag agttggaagc agtggataat tgaacgtgaa cgtgtctgat
      901 gttggacaag ttagctctgt tcaacaattc tgggtaacga ccttgtaagt tttcaatggc
      961 ttcaactgga ccgtggacta aaccacaaca agcgtacaat ttgattctga tggtgaaagt
     1021 gtgccagaaa gaacctaaac cggcaacaac ttcttcttcc ttgtatggag gttctctacc
     1081 gttacccttg gtgaacatct tcaagaaacc accgtaagat ctttccaaga cacccttcat
     1141 agcttcgtaa ccaccgtgag ccaacaaacc acctaataga ccgtttctgg cagcgaaacc
     1201 atgttgaact ctcttaacca taccaccgta ttgagcagac atcaaaccac aagcttgggt
     1261 acaagcgata cccaaagcat cttccatgga gtctggagtc aaacccaaca acttaccggt
     1321 agccaaagca ccagctggag caccgtaaac ggcaccacaa tgccaaccgt tgttcaacaa
     1381 gtcagaaccg tagatggcct taccgattct tggaccagat tcgaaaccga caatggcagc
     1441 taagataaca tcgataccag agatggtctt accttgttca gccaagactt cagaagcagc
     1501 gaaaacagct ggtagaacaa tggaagcaga gtgcaatgga gcttcagagt ggtagtcatc
     1561 caattcagtg gcttgaataa aggcagagtt ggtcatggca gcagcaactg gacccaattt
     1621 ttgaccgtaa ccgatgacac gacaagcacc tggaggttcg aaggacatgg tagcttgaac
     1681 gtacttttca gaccatggaa cacgagcacc aacccaagca caggcaatac cgtccaaaat
     1741 caagtacttg gctctttcca agacatcaga tggaatgtcg tcagtagcca agttggaagc
     1801 ccagtgacag atttcggaag tgacaccaga cttggcattg gagtcagcag attgcttggt
     1861 cattttgtaa ttaaaactta gatcagtttg ctatgctctc tttctaattg accaatgctg
     1921 agaagaagaa tgactaaaca agaggaacaa attaaaactt ggaaaacggg agtgctttta
     1981 ttaacttaaa tatcaatggg aggtcatcga aagagaaaaa atcccaaaaa aaattttcaa
     2041 gaaaaagaaa tgtgataaaa attttttttt ttgccttttc cgacaaagaa aaagaaacga
     2101 aatagtcttt tgttactttt atacagtttc atgcgggtaa tggagccgca cgaaagagac
     2161 aaagagggga aacatggtat gaggtgtgtg ggtgtggctt ggctcgtgcg gcgatgcgga
     2221 gagtgcgaga aaatccggaa gagtaaaaag gagtacagtg cgtttttgaa gcatgcactg
     2281 ctaagatttg tgttcgtttg ggtgcagtcg gttgtgtaca tcgatccgcc ccacttcaga
     2341 agttcctata ctttctagag aataggaact tcactataga gtcgaataag ggcgacaccc
     2401 cctaattagc ccgggcgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     2461 ctggcagttc cctactctcg catggggagt ccccacacta ccatcggcgc tacggcgttt
     2521 cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaaca
     2581 aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc agaattggtt
     2641 aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2701 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga
     2761 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     2821 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     2881 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2941 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcca cttccgacca
     3001 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     3061 aaacagcgtt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     3121 tggcagtgtt cctgcgccgg ttgcactcga ttcctgtttg taattgtcct tttaacagcg
     3181 atcgcgtatt tcgcctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     3241 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     3301 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     3361 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     3421 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     3481 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcaatttc
     3541 atttgatgct cgatgagttt ttctaagcag agcattacgc tgacttgacg ggacggcgca
     3601 agctcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
     3661 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3721 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3781 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3841 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
     3901 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3961 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4021 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4081 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4141 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4201 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4261 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4321 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4381 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4441 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     4501 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     4561 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     4621 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     4681 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     4741 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     4801 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     4861 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     4921 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     4981 ttattat
//
