LOCUS       Exported                5057 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-40, complete sequence.
ACCESSION   KM216416
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     pUMRI plasmids for metabolic pathway assembly
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5057)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUL-2014) Department of Chemical and Biological 
            Engineering, Zhejiang University, Institute of Bioengineering, 34 
            TianMuShan Road, Hangzhou, Zhejiang 310028, China
REFERENCE   3  (bases 1 to 5057)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Artificial Sequence v. Artificial Sequence 
            Sequencing Technology :: Artificial Sequence
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5057
                     /organism="Cloning vector pMRI-40"
                     /mol_type="other DNA"
                     /db_xref="taxon:1563409"
     misc_feature    complement(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..1217)
                     /regulatory_class="terminator"
                     /note="PGK1 terminator from Saccharomyces 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      complement(1293..1811)
                     /regulatory_class="promoter"
                     /note="GAL2 promoter from Saccharomyces cerevisiae"
     regulatory      1812..2360
                     /regulatory_class="promoter"
                     /note="GAL7 promoter from Saccharomyces cerevisiae"
     promoter        2368..2386
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2404..2433
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2459..2791
                     /regulatory_class="terminator"
                     /note="TPS1 terminator from Saccharomyces cerevisiae"
     misc_feature    complement(2813..2846)
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2813..2846)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2892..4248
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2892..3235
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3236..4045
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to kanamycin and geneticin"
                     /protein_id="AIU45944.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      4051..4248
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(4381..4969)
                     /direction=LEFT
                     /label=ColE1 pBR322
                     /note="ColE1 pBR322"