LOCUS       40924_32275        4704 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-34, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4704)
  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  JOURNAL   Unpublished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 4704)
  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 4704)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4704)
  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            "Unpublished"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..4704
                     /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..1029
                     /label=DPP1 homologous arm
                     /note="DPP1 homologous arm"
     terminator      complement(1064..1229)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1377..1400)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        1447..2111
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        2122..2140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2158..2187
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      2217..2406
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2419..2437)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2460..2493
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2460..2493)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2539..3895
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(4028..4616)
                     /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 gtcaaccgat
      121 cgacaaatta tttctaaaat acaaaaaagt tagtcgtgag gttatccaac gttgggtttc
      181 tatctatata taaaagctaa tcaccaaaca attgagaatc aacgccgata acatgatgta
      241 caattatagc agtatacgcc aggcgtgctg catttattct ttaccctgtc atcgttgacg
      301 aatacttttt tttaaaggga tcgcactcta accacctgaa aggtactatc tgaaaaagtg
      361 ttgactaaaa acagaagcgc cttaataata gtgcactact ggaaaactat atttaatgta
      421 aaaaagaaac tacattgaat gaaatcgaaa agtataattt ctctccgtac ctgtgactga
      481 tacaatgtaa cgatgtagac taagaataag attctcactt aaaagctgct atttcaactg
      541 attatctgca gagtatgcga tcacaagaca gatcacaact gtgtttatcg gacataatgc
      601 ggccataaag gcctgatggg tgactgcttc ctccagggtg acatctgaat cgtccatcaa
      661 tggtttgaac ggaagaggat catcaatggg tgggaagatt cttctgtaga aaaagtgtgc
      721 cattatataa cccaacatag accctaaaat tacatcgacg aaatgatgtc tgtaatcttg
      781 agttctggat agagcaatta gtgcagctcc taacagtggt agaaaggcca ccatttttct
      841 ccataaaggc atcaacggtg attcagtcaa aagttgccca catagccaga agtacaaata
      901 acccagtcct gcaaagcttt cacttgaatg acctgacgga gttgtcctaa acccatccaa
      961 cagacgttcg tgattcttag tcgtacacac atcttttgca gtaaataaag tgtccaatgg
     1021 caagccttct gtaggcctct tcgctattac gccagctgaa ttggagcgac ctcatgctat
     1081 acctgagaaa gcaacctgac ctacaggaaa gagttactca agaataagaa ttttcgtttt
     1141 aaaacctaag agtcacttta aaatttgtat acacttattt tttttataac ttatttaata
     1201 ataaaaatca taaatcataa gaaattcgct tatttagaag tgtcaacaac gtatctacca
     1261 acgatttgac ccttttccat cttttcgtaa atttctggca aggtagacaa gccgacaacc
     1321 ttgattggag acttgaccaa acctctggcg aagaattgtt aattaagagc tcagatctta
     1381 tcgtcgtcat ccttgtaatc catcgatact agtgcggccg ccctttagtg agggttgaat
     1441 tcgaattttc aaaaattctt actttttttt tggatggacg caaagaagtt taataatcat
     1501 attacatggc attaccacca tatacatatc catatacata tccatatcta atcttactta
     1561 tatgttgtgg aaatgtaaag agccccatta tcttagccta aaaaaacctt ctctttggaa
     1621 ctttcagtaa tacgcttaac tgctcattgc tatattgaag tacggattag aagccgccga
     1681 gcgggtgaca gccctccgaa ggaagactct cctccgtgcg tcctcgtctt caccggtcgc
     1741 gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg aacaataaag attctacaat
     1801 actagctttt atggttatga agaggaaaaa ttggcagtaa cctggcccca caaaccttca
     1861 aatgaacgaa tcaaattaac aaccatagga tgataatgcg attagttttt tagccttatt
     1921 tctggggtaa ttaatcagcg aagcgatgat ttttgatcta ttaacagata tataaatgca
     1981 aaaactgcat aaccacttta actaatactt tcaacatttt cggtttgtat tacttcttat
     2041 tcaaatgtaa taaaagtatc aacaaaaaat tgttaatata cctctatact ttaacgtcaa
     2101 ggagaaaaaa ccccggatcc gtaatacgac tcactatagg gcccgggcgt cgacatggaa
     2161 cagaagttga tttccgaaga agacctcgag taagcttggt accgcggcta gctaagatcc
     2221 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     2281 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     2341 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     2401 tcgaagatcc caattcgccc tatagtgagt cgtattacgc gcgctcgaca acccttaata
     2461 taacttcgta taatgtatgc tatacgaagt tattaggtct agtagcttgc ctcgtccccg
     2521 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     2581 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     2641 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     2701 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     2761 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     2821 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     2881 ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg
     2941 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3001 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3061 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3121 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3181 ccggcaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3241 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3301 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3361 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3421 tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3481 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3541 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3601 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3661 agtttcattt gatgctcgat gagtttttct aatcagtact gacaataaaa agattcttgt
     3721 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     3781 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     3841 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc
     3901 gattcgatac taacgccgcc atccagtgtc gaaaacgaac agaatcaggg gataacgcag
     3961 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4021 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4081 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4141 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4201 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4261 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4321 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4381 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4441 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4501 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4561 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4621 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4681 ttttggtcat gagattatca aaaa
//
