LOCUS       40924_32255        4660 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-28, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4660)
  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  JOURNAL   Unpublished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 4660)
  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 4660)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4660)
  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            "Unpublished"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..4660
                     /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..1618)
                     /label=HXT1 promoter derived from S. cerevisiae
                     /note="HXT1 promoter derived from S. cerevisiae"
                     /regulatory_class="promoter"
     promoter        1686..2093
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2114..2132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2173..2362
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2375..2393)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2416..2449
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2416..2449)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2495..3851
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(3984..4572)
                     /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 cgattttacg tatatcaact agttgacgat tatgatatct
      541 ttatagattt taaggtgggg aaagaacatg agaccccaga tggaattgat tatggggaca
      601 ttgttgcctt tatatataat ttcaatatac taattcaaat gattaaaaac gtgaggggga
      661 cacgcaactt cgggtgttaa gaaatatttt gctacattag ataatggtgg agtttcctgg
      721 cttgtcggat aaaagccatc aaatgtcgca gcagctcatg tttacgtttg ctgtcttctg
      781 cccacgtcat atgagtggta ttcttctatc agcacttgat gaatattctt tttctcatat
      841 atctgaaaga caaaagatcg gcacggcaat gccctgcagc atttcttcct agtttttccg
      901 aatttccatt acgtattgga tcttgtgcgc atatttgtca gtccttcacg gaaaaaaaaa
      961 aagagcactg ggtcacttcg gaaaaacttt tgactcaatg caacagtgtc ataatccttt
     1021 gcgctgtctc tttgaagaaa aatcaggagt gcaagatatc gattaattcc ttggaagtta
     1081 tgatggttag tcttagttta actctcttga agaagggttt tttcagttgg tcaacactct
     1141 ttagaggtaa aaaaaaaaaa aaaaaaaaaa aaaagagaat tcttcatgta atttaccatg
     1201 attctacgtt tttgcaagca aaaatgaaga taatccgagc gcatgcgcaa gtagtccctg
     1261 ccatgccgct tcggaaaaac tttcgaaacc aatactcctt acatccgcct tgaatttagg
     1321 acctacaatt gttgtctctt ctgcttgaac caataagccc tagaaacctt acaccctaat
     1381 ttgcacaaga aaactacgaa aaaatttttt cctctttttt ccgcagctcc aatctcgccc
     1441 tgctattttt ttggttcgga gaaaaatagt cggtcgatat gaattgaagt ttcattgtgg
     1501 ccactcgttt aggaatggaa taacctccgc gaatggaaaa aaagaactcg gaatatcaaa
     1561 aattccggct caaaggaaaa atttgcttca ggagggggga attatattcc agatgagacc
     1621 tgcatacccc tcatttccac gaccgcgaat ccttacatca cacccaatcc cccacaagtg
     1681 atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc
     1741 tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc
     1801 cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa
     1861 gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc
     1921 tttttcttga aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt
     1981 taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac
     2041 aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct aagttttaat
     2101 tacaaaggat ccgtaatacg actcactata gggcccgggc gtcgagtaag cttggtaccg
     2161 cggctagcta agatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     2221 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     2281 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     2341 gaaggttttg ggacgctcga agatcccaat tcgccctata gtgagtcgta ttacgcgcgc
     2401 tcgacaaccc ttaatataac ttcgtataat gtatgctata cgaagttatt aggtctagta
     2461 gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct
     2521 tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc
     2581 catacatccc catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag
     2641 caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc
     2701 gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact
     2761 gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac
     2821 atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc gcgattaaat
     2881 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2941 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     3001 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     3061 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     3121 ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata tcctgattca
     3181 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     3241 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     3301 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3361 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     3421 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3481 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3541 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3601 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agtactgaca
     3661 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     3721 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     3781 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     3841 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa acgaacagaa
     3901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4381 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
//
