LOCUS       Exported                5948 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUMRI-10, complete sequence.
ACCESSION   KM216412
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5948)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     pUMRI plasmids for metabolic pathway assembly
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5948)
  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 5948)
  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..5948
                     /organism="Cloning vector pUMRI-10"
                     /mol_type="other DNA"
                     /db_xref="taxon:1563405"
     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    105..1142
                     /label=HO homologous arm
                     /note="HO homologous arm"
     CDS             complement(1078..1089)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      complement(1143..1438)
                     /regulatory_class="terminator"
                     /note="ADH1 terminator from Saccharomyces cerevisiae"
     terminator      1153..1318
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1466..1489)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1536..2200
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     regulatory      1540..2199
                     /regulatory_class="promoter"
                     /note="GAL1 and GAL10 bidirectional promoters from 
                     Saccharomyces cerevisiae"
     protein_bind    1752..1869
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        2211..2229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2247..2276
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2303..2480
                     /regulatory_class="terminator"
                     /note="CYC1 terminator from Saccharomyces cerevisiae"
     terminator      2306..2495
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2508..2526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(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)"
     promoter        2628..2971
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2972..3781
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to kanamycin and geneticin"
                     /protein_id="AIU45936.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      3787..3984
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        4084..4299
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4300..5103
                     /codon_start=1
                     /product="orotidine-5-phosphate (OMP) decarboxylase"
                     /label=orotidine-5-phosphate
                     /note="URA3"
                     /protein_id="AIU45937.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5272..5860)
                     /direction=LEFT
                     /label=ColE1 pBR322
                     /note="ColE1 pBR322"