LOCUS       Exported                5028 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-35, complete sequence.
ACCESSION   KJ502284
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5028)
  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  JOURNAL   Unpublished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 5028)
  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 5028)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5028
                     /organism="Cloning vector pMRI-35"
                     /mol_type="other DNA"
                     /db_xref="taxon:1495078"
     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..917
                     /label=LPP1 homologous arm
                     /note="LPP1 homologous arm"
     regulatory      complement(942..1843)
                     /regulatory_class="promoter"
                     /note="HXT7 promoter derived from S. cerevisiae"
     regulatory      complement(942..1217)
                     /regulatory_class="terminator"
                     /note="PGK1 terminator derived from S. cerevisiae"
     terminator      947..1217
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             complement(1236..1259)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1866..2331
                     /regulatory_class="promoter"
                     /note="TEF1 promoter derived from S. cerevisiae"
     promoter        1911..2318
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2339..2357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2375..2404
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2430..2762
                     /regulatory_class="terminator"
                     /note="TPS1 terminator derived from S. cerevisiae"
     misc_feature    2784..2817
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2784..2817)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2863..4219
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2863..3206
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3207..4016
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to kanamycin and geneticin"
                     /protein_id="AHY82610.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      4022..4219
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ColE1 pBR322
                     /note="ColE1 pBR322"