LOCUS       40924_32250        3784 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-21, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3784)
  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  JOURNAL   Unpublished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 3784)
  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 3784)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3784)
  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            "Unpublished"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..3784
                     /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)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        527..1191
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        1202..1220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1238..1267
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      1297..1486
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1499..1517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1540..1573
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(1540..1573)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            1619..2975
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(3108..3696)
                     /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 ccctttagtg agggttgaat tcgaattttc aaaaattctt
      541 actttttttt tggatggacg caaagaagtt taataatcat attacatggc attaccacca
      601 tatacatatc catatacata tccatatcta atcttactta tatgttgtgg aaatgtaaag
      661 agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac
      721 tgctcattgc tatattgaag tacggattag aagccgccga gcgggtgaca gccctccgaa
      781 ggaagactct cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg
      841 cctcgcgccg cactgctccg aacaataaag attctacaat actagctttt atggttatga
      901 agaggaaaaa ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac
      961 aaccatagga tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg
     1021 aagcgatgat ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta
     1081 actaatactt tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc
     1141 aacaaaaaat tgttaatata cctctatact ttaacgtcaa ggagaaaaaa ccccggatcc
     1201 gtaatacgac tcactatagg gcccgggcgt cgacatggaa cagaagttga tttccgaaga
     1261 agacctcgag taagcttggt accgcggcta gctaagatcc gctctaaccg aaaaggaagg
     1321 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     1381 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     1441 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaagatcc caattcgccc
     1501 tatagtgagt cgtattacgc gcgctcgaca acccttaata taacttcgta taatgtatgc
     1561 tatacgaagt tattaggtct agtagcttgc ctcgtccccg ccgggtcacc cggccagcga
     1621 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
     1681 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
     1741 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca
     1801 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     1861 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     1921 gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact
     1981 cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2041 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     2101 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2161 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2221 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag
     2281 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2341 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2401 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2461 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc
     2521 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2581 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2641 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2701 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2761 gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg
     2821 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     2881 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     2941 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc
     3001 atccagtgtc gaaaacgaac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3541 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3781 aaaa
//
