LOCUS       40924_32265        4793 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-31, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  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  JOURNAL   Unpublished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 4793)
  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 4793)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4793)
  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            "Unpublished"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..4793
                     /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)."
     CDS             complement(1078..1089)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     terminator      complement(1153..1318)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1466..1489)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        1536..2200
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        2211..2229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2247..2276
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      2306..2495
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2508..2526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2549..2582
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2549..2582)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2628..3984
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(4117..4705)
                     /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 gtatcgagat
      121 cacttttcgt gatccgctaa tcagcgacgg tcacattagg tttgccaagt cagggtatga
      181 accatacgat cagttttcgt gaacctggta cgtatattgt ggcgtttgtg tatattttca
      241 ttctttgaca acaatcaata ccaacctcaa ataggaaaag taataagttt ggcgttacac
      301 cccaaaagac gccaaacgga tcgaacttac tcaatagcaa ttagcgagac aaaacctacg
      361 ttaagacctg taaccgattt atcaaagcac tctgcggttc tttcttggga atattacctg
      421 gacattttgt gccctcaaga aacgaggctc tacgagcctg ttggagcccc tcagacatta
      481 gccgccacga atcaaacttt ttacgcgatt cggcccatga ggcccgcgga cagcatcaaa
      541 ctgtaagatt ccgccacatt ttatacactc tggtccttta actggcaaac cttcgggcgt
      601 aatgcccaat ttttcgcctt tgtcttttgc ctttttcact tcacgtgctt ctggtacata
      661 cttgcaattt atacagtgat gaccgctgaa tttgtatctt ccatagcatc tagcacatac
      721 tcgattttta ccactccaat ctttataaaa atacttgatt ccctttctgg gacaagcaac
      781 acagtgtttt agattctttt tttgtgatat tttaagctgt tctcccacac agcagcctcg
      841 acatgatttc acttctattt tgttgccaag caagaaattt ttatggcctt ctatcgtaag
      901 cccatataca gtactctcac cctggaaatc atccgtgaag ctgaaatata cgggttccct
      961 ttttataatt ggcggaactt ctcttgtttt gtgaccactt cgacaatatg acaaaacatt
     1021 ctgtgaagtt gttcccccag caacattaca gtcgtatgta aattgacatt ggacttttct
     1081 tccttcaatg atttcctccc tagctgacct ggtcgtcttg taggcctctt cgctattacg
     1141 ccagctgaat tggagcgacc tcatgctata cctgagaaag caacctgacc tacaggaaag
     1201 agttactcaa gaataagaat tttcgtttta aaacctaaga gtcactttaa aatttgtata
     1261 cacttatttt ttttataact tatttaataa taaaaatcat aaatcataag aaattcgctt
     1321 atttagaagt gtcaacaacg tatctaccaa cgatttgacc cttttccatc ttttcgtaaa
     1381 tttctggcaa ggtagacaag ccgacaacct tgattggaga cttgaccaaa cctctggcga
     1441 agaattgtta attaagagct cagatcttat cgtcgtcatc cttgtaatcc atcgatacta
     1501 gtgcggccgc cctttagtga gggttgaatt cgaattttca aaaattctta cttttttttt
     1561 ggatggacgc aaagaagttt aataatcata ttacatggca ttaccaccat atacatatcc
     1621 atatacatat ccatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat
     1681 cttagcctaa aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct
     1741 atattgaagt acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc
     1801 ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc
     1861 actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat
     1921 tggcagtaac ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat
     1981 gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt
     2041 tttgatctat taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt
     2101 caacattttc ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt
     2161 gttaatatac ctctatactt taacgtcaag gagaaaaaac cccggatccg taatacgact
     2221 cactataggg cccgggcgtc gacatggaac agaagttgat ttccgaagaa gacctcgagt
     2281 aagcttggta ccgcggctag ctaagatccg ctctaaccga aaaggaagga gttagacaac
     2341 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     2401 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     2461 aaaccttgct tgagaaggtt ttgggacgct cgaagatccc aattcgccct atagtgagtc
     2521 gtattacgcg cgctcgacaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     2581 attaggtcta gtagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
     2641 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
     2701 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
     2761 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
     2821 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
     2881 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
     2941 gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag
     3001 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     3061 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     3121 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     3181 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     3241 tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga
     3301 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3361 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3421 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3481 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
     3541 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3601 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3661 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3721 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3781 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     3841 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     3901 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     3961 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
     4021 aaaacgaaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaa
//
