LOCUS       40924_32270        4902 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-32, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4902)
  AUTHORS   Xie W, Yu H.
  TITLE     Developing a sequential repressing-inducing control strategy for 
            improving carotenoids production in Saccharomyces cerevisiae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4902)
  AUTHORS   Xie W, Yu H.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2014) Department of Chemical and Biological 
            Engineering, Zhejiang University, Institute of Bioengineering, 34 
            TianMuShan Road, Hangzhou, Zhejiang 310028, China
REFERENCE   3  (bases 1 to 4902)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4902)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2014) Department of Chemical and Biological Engineering, 
            Zhejiang University, Institute of Bioengineering, 34 TianMuShan 
            Road, Hangzhou, Zhejiang 310028, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..55
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    113..1227
                     /label=Ty4 homologous arm
                     /note="Ty4 homologous arm"
     terminator      complement(1262..1427)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1575..1598)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        1645..2309
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        2320..2338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2356..2385
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      2415..2604
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2617..2635)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2658..2691
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2658..2691)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2737..4093
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(4226..4814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggatcttcga gaacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt
       61 gatatcagat ccacggatca ctaaagggaa caaaagctgg agctggcctt gtgcaaaaat
      121 gcgttttact tcctttacaa tctcaatgcc gataaataag cgcaatgaag attactcagc
      181 aactgtatgt tggctcatgc gaaatatgaa tgtttcattc gagattactt atcttgttta
      241 acgcttgaac agcgtactgt aacttcttgc tacagaatga tggaccccat tataatacta
      301 ccgccttgcg aatgcgcaat ttattatgat catgaaccct gagaaatcaa atgaatattt
      361 aaagtgtatg caaaatggac aagaacatta tggtaacatt gtctacgaca gaatttattt
      421 tctgacttta atagtgacgg ttatacgaaa ttaatgattc ttcgagaggc ctttatggcc
      481 cgtaatacaa cagtgagcta ttaaaagagc ttaagtatta cagagtaata tcaatcatca
      541 atcttggatc taaagtaatc gtcgagacat ttatacactt aagtgattcc tctttacaag
      601 cggagcttac tctatatatt cattcttgtc agtttcataa cagttgtttc atacgatttc
      661 cacatgtgtc tcatatatat tttatgttta ggttaataac tttggtaatg ctaaattatc
      721 ttaatttgac gtacacaaat tcttcctact gttatatatg ttcgaattat cttttaaatt
      781 gttgctaaaa aagttagagt tatcaccaaa tcagttgagc agatcaccat tattcttatg
      841 taaatacttg ttaaaagaag tagctttcgc aaatacacca atgcattcgt tgataattta
      901 cgcagtaacc tcttacagaa cgcctacata caacttaaac tcgtaatgtt caggcttata
      961 atgtcagtat gcattgttac gcttagtata gttattaaac tacggaccaa attatacttt
     1021 aagcagtagt taaagaaaga gattaaataa agaatgattt acaatctagt cgcaaaaaca
     1081 agtacagtgc tgacgtccca tcttttaatg cctactcttt tgatgttcac ttacatgtta
     1141 caatgaagcc aaatcacaaa cattcaaata tattccatgg cctcttagtt tggcaaccca
     1201 agactcggca taccatattg gtaacgctgt aggcctcttc gctattacgc cagctgaatt
     1261 ggagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
     1321 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
     1381 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg
     1441 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag
     1501 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa
     1561 ttaagagctc agatcttatc gtcgtcatcc ttgtaatcca tcgatactag tgcggccgcc
     1621 ctttagtgag ggttgaattc gaattttcaa aaattcttac tttttttttg gatggacgca
     1681 aagaagttta ataatcatat tacatggcat taccaccata tacatatcca tatacatatc
     1741 catatctaat cttacttata tgttgtggaa atgtaaagag ccccattatc ttagcctaaa
     1801 aaaaccttct ctttggaact ttcagtaata cgcttaactg ctcattgcta tattgaagta
     1861 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
     1921 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
     1981 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
     2041 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
     2101 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
     2161 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
     2221 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
     2281 tctatacttt aacgtcaagg agaaaaaacc ccggatccgt aatacgactc actatagggc
     2341 ccgggcgtcg acatggaaca gaagttgatt tccgaagaag acctcgagta agcttggtac
     2401 cgcggctagc taagatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     2461 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     2521 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     2581 gagaaggttt tgggacgctc gaagatccca attcgcccta tagtgagtcg tattacgcgc
     2641 gctcgacaac ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtctag
     2701 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     2761 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     2821 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     2881 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     2941 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     3001 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     3061 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
     3121 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     3181 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
     3241 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     3301 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     3361 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
     3421 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     3481 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     3541 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     3601 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
     3661 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     3721 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     3781 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     3841 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga
     3901 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     3961 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     4021 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     4081 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtgtcga aaacgaacag
     4141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aa
//
