LOCUS       Exported                4902 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-32, complete sequence.
ACCESSION   KJ502282
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4902)
  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  JOURNAL   Unpublished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 4902)
  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 4902)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4902
                     /organism="Cloning vector pMRI-32"
                     /mol_type="other DNA"
                     /db_xref="taxon:1495076"
     misc_feature    22..55
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(22..55)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    113..1227
                     /label=Ty4 homologous arm
                     /note="Ty4 homologous arm"
     regulatory      complement(1254..1555)
                     /regulatory_class="terminator"
                     /note="ADH1 terminator derived from S. cerevisiae"
     terminator      1262..1427
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1575..1598)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1644..2309
                     /regulatory_class="promoter"
                     /note="GAL1 and GAL10 bidirectional promoters derived from 
                     S. cerevisiae"
     promoter        1645..2309
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    1861..1978
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        2320..2338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2356..2385
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2415..2604
                     /regulatory_class="terminator"
                     /note="CYC1 terminator derived from S. cerevisiae"
     terminator      2415..2604
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2617..2635)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2658..2691
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2658..2691)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2737..4093
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2737..3080
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3081..3890
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to kanamycin and geneticin"
                     /protein_id="AHY82608.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      3896..4093
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(4226..4814)
                     /direction=LEFT
                     /label=ColE1 pBR322
                     /note="ColE1 pBR322"