LOCUS       40924_32260        3876 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-29, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3876)
  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  JOURNAL   Unpublished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 3876)
  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 3876)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3876)
  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            "Unpublished"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..3876
                     /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)."
     terminator      complement(144..309)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(457..480)
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     regulatory      complement(502..834)
                     /label=BTS1 promoter derived from S. cerevisiae
                     /note="BTS1 promoter derived from S. cerevisiae"
                     /regulatory_class="promoter"
     promoter        902..1309
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1330..1348
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1389..1578
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1591..1609)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1632..1665
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(1632..1665)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            1711..3067
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(3200..3788)
                     /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 gtaggcctct
      121 tcgctattac gccagctgaa ttggagcgac ctcatgctat acctgagaaa gcaacctgac
      181 ctacaggaaa gagttactca agaataagaa ttttcgtttt aaaacctaag agtcacttta
      241 aaatttgtat acacttattt tttttataac ttatttaata ataaaaatca taaatcataa
      301 gaaattcgct tatttagaag tgtcaacaac gtatctacca acgatttgac ccttttccat
      361 cttttcgtaa atttctggca aggtagacaa gccgacaacc ttgattggag acttgaccaa
      421 acctctggcg aagaattgtt aattaagagc tcagatctta tcgtcgtcat ccttgtaatc
      481 catcgatact agtgcggccg ctgattttcc agactcgtaa acgctttgtt ctctttctat
      541 tagtagcttc tttgaaaatg agccttacat ggttttgtaa gcacatctgt cttgactttt
      601 tttttgcttc aaaaaaacgt caaagaaggc aaggcgaagc aaagaaaaat ggtagtaagc
      661 atacaatgat cagcataaca tatgcaaaaa cacgtaattt ctgctatgct ggaagcggtg
      721 aacgttggat accactataa taatagaagt ttacctttca gtatgaaaat gcgggcttgt
      781 cctatatcta tatgtaatat tatggagtga gtagagatgg cggctataca tcgtcctgca
      841 tacccctcat ttccacgacc gcgaatcctt acatcacacc caatccccca caagtgatcc
      901 cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
      961 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     1021 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     1081 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     1141 tcttgaaaat tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag
     1201 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
     1261 ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca
     1321 aaggatccgt aatacgactc actatagggc ccgggcgtcg agtaagcttg gtaccgcggc
     1381 tagctaagat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1441 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1501 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     1561 gttttgggac gctcgaagat cccaattcgc cctatagtga gtcgtattac gcgcgctcga
     1621 caacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt ctagtagctt
     1681 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1741 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1801 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1861 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1921 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1981 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     2041 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
     2101 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2161 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2221 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2281 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2341 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2401 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2461 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2521 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2581 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2641 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2701 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2761 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2821 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     2881 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     2941 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     3001 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     3061 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga acagaatcag
     3121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3601 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3841 cacgttaagg gattttggtc atgagattat caaaaa
//
