LOCUS       sequence-254054-        5100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 3. CAD transcription unit with the RPS3 
            promoter and CYC1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5100)
  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 5100)
  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-Prps3-CAD-Tcyc1
FEATURES             Location/Qualifiers
     source          1..5100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    486..519
                     /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     741..759
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(2893..2926)
                     /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(3133..3152)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3219..3310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3311..4120
                     /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      4242..4830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4731..4750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4984..5001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     join(5099..5100,1..16)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(5100,1..16)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taaaacgacg gccagtctta agctcgggcc ccctgggcgg ttctgataac gagtaatcgt
       61 taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt tttatggggg gagtttaggg
      121 aaagagcatt tgtcagaata tttaagggcg cctgtcactt tgcttgatat atgagaatta
      181 tttaacctta taaatgagaa aaaagcaacg cactttaaat aagatacgtt gctttttcga
      241 ttgatgaaca cctataatta aactattcat ctattattta tgattttttg tatatacaat
      301 atttctagtt tgttaaagag aattaagaaa ataaatctcg aaaataataa agggaaaatc
      361 agtttttgat atcaaaatta tacatgtcaa cgataataca aaatataata caaactataa
      421 gatgttatca gtatttatta tgcatttaga ataaattttg tgtcgccctt attcgactca
      481 ctatagaagt tcctattctc tagaaagtat aggaacttca cttcattttc aacaggaggc
      541 ggatggatat actgtggtct ggaagatgcc ggaaagcgtg aggtgttaca tgcgtacacg
      601 cgtttgtaca gaaaaaaaag aaaaatttga aatataaata acgttcttaa tactaacata
      661 actataaaaa aataaatagg gacctagact tcaggttgtc taactccttc cttttcggtt
      721 agagcggatg tggggggagg gcgtgaatgt aagcgtgaca taactaatta catgatatcg
      781 acaaaggaaa aggggcctgt ttacttgata cccaatggag atttgactgg acagttcaac
      841 aattccaata atggagagat atcggtcaat ctgtccaaac ccaaaaccaa atcttcgatt
      901 tccttgactc tggattcgtc agtgacggaa ccagccaaag ttctgtattt gtgcaagatt
      961 ctttcgtttg gcattggttc cttgacacct aatggctttt caacggattc agtgatggaa
     1021 gaaccgtcgt tgaattcaat tctgactcta ccagcagaca aacagttacc atcttggtca
     1081 aattcttcag attgagaaga ggtgaccttt ctagccaaat cccagacttc tggtctttcc
     1141 aagttgtcat cgaattcgga gaattgagac aacaaacatt gttggtcaac caattggaca
     1201 gccaaaatgt aagcaacaga catttgacca gcaatggagg agattggtct ttcttctgga
     1261 atccaaccac agtgagagtt ggaagcagtg gataattgaa cgtgaacgtg tctgatgttg
     1321 gacaagttag ctctgttcaa caattctggg taacgacctt gtaagttttc aatggcttca
     1381 actggaccgt ggactaaacc acaacaagcg tacaatttga ttctgatggt gaaagtgtgc
     1441 cagaaagaac ctaaaccggc aacaacttct tcttccttgt atggaggttc tctaccgtta
     1501 cccttggtga acatcttcaa gaaaccaccg taagatcttt ccaagacacc cttcatagct
     1561 tcgtaaccac cgtgagccaa caaaccacct aatagaccgt ttctggcagc gaaaccatgt
     1621 tgaactctct taaccatacc accgtattga gcagacatca aaccacaagc ttgggtacaa
     1681 gcgataccca aagcatcttc catggagtct ggagtcaaac ccaacaactt accggtagcc
     1741 aaagcaccag ctggagcacc gtaaacggca ccacaatgcc aaccgttgtt caacaagtca
     1801 gaaccgtaga tggccttacc gattcttgga ccagattcga aaccgacaat ggcagctaag
     1861 ataacatcga taccagagat ggtcttacct tgttcagcca agacttcaga agcagcgaaa
     1921 acagctggta gaacaatgga agcagagtgc aatggagctt cagagtggta gtcatccaat
     1981 tcagtggctt gaataaaggc agagttggtc atggcagcag caactggacc caatttttga
     2041 ccgtaaccga tgacacgaca agcacctgga ggttcgaagg acatggtagc ttgaacgtac
     2101 ttttcagacc atggaacacg agcaccaacc caagcacagg caataccgtc caaaatcaag
     2161 tacttggctc tttccaagac atcagatgga atgtcgtcag tagccaagtt ggaagcccag
     2221 tgacagattt cggaagtgac accagacttg gcattggagt cagcagattg cttggtcatt
     2281 tttgtagttt gtttgctgtt ttattttcag ttatcaaaac aatgttcaaa aactaatcta
     2341 acaatatcta caaactagaa attgcttaaa atgtatgaaa aatgctgttg tatttcaaag
     2401 agctgcattg atttggaaaa tttaccactg cccatgggag aaaggctcgc ggcgcctcat
     2461 ctcaagaaaa gaaaatgttt ttttcataaa aaaaacaaac tttacagcaa gaatattata
     2521 caggaaaggt atggatgtta cggaaacaaa aaacagtctc atttaatcat gaggaattag
     2581 attattccaa aggaacatga aaagaagtga aatcaagaaa aaaaaacaag agaatttacc
     2641 ggcggccaaa aatactaacg taatcccgtt gacgtttacg catgcagtgt tgtaggctca
     2701 ccagcaatta tgttagttga gatgtaactg cttcgcgtga acactacgta aatgttattg
     2761 tgtggaactg acgtggacct tttctgtgtt gcttttagta taacgatatc catcctttta
     2821 cttggattgc actgctaaga tttgtgttcg tttgggtgca gtcggttgtg tacatcgatc
     2881 cgccccactt cagaagttcc tatactttct agagaatagg aacttcacta tagagtcgaa
     2941 taagggcgac accccctaat tagcccgggc gaaaggccca gtctttcgac tgagcctttc
     3001 gttttatttg atgcctggca gttccctact ctcgcatggg gagtccccac actaccatcg
     3061 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
     3121 ccgccaggca aacaaggggt gttatgagcc atattcaggt ataaatgggc tcgcgataat
     3181 gttcagaatt ggttaattgg ttgtaacact gacccctatt tgtttatttt tctaaataca
     3241 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3301 aaggaagaat atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     3361 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3421 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3481 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3541 gccacttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3601 tgcgatcccc ggaaaaacag cgttccaggt attagaagaa tatcctgatt caggtgaaaa
     3661 tattgttgat gcgctggcag tgttcctgcg ccggttgcac tcgattcctg tttgtaattg
     3721 tccttttaac agcgatcgcg tatttcgcct cgctcaggcg caatcacgaa tgaataacgg
     3781 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3841 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3901 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3961 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     4021 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     4081 taaattgcaa tttcatttga tgctcgatga gtttttctaa gcagagcatt acgctgactt
     4141 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc
     4201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4381 aatactgttc ttctagtgta gccgtagtta gcccaccact tcaagaactc tgtagcaccg
     4441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4981 cgcagcgagt cagtgagcga ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa
     5041 ctaagcagaa ggcccctgac ggatggcctt tttgcgtttc tacaaactct ttctgtgttg
//
