LOCUS       Exported                6866 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUMRI-12, complete sequence.
ACCESSION   KM216414
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6866)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     pUMRI plasmids for metabolic pathway assembly
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6866)
  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 6866)
  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..6866
                     /organism="Cloning vector pUMRI-12"
                     /mol_type="other DNA"
                     /db_xref="taxon:1563407"
     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    118..795
                     /label=ERG9 upstream region
                     /note="ERG9 upstream region"
     misc_feature    complement(796..1499)
                     /label=ERG9 region
                     /note="ERG9 region"
     regulatory      complement(1497..2613)
                     /regulatory_class="promoter"
                     /note="HXT1 promoter from Saccharomyces cerevisiae"
     regulatory      2614..3121
                     /regulatory_class="promoter"
                     /note="GAL7 promoter from Saccharomyces cerevisiae"
     promoter        3129..3147
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3165..3194
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3224..3413
                     /regulatory_class="terminator"
                     /note="CYC1 terminator from Saccharomyces cerevisiae"
     terminator      3224..3413
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(3426..3444)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(3467..3500)
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(3467..3500)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            3546..4902
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        3546..3889
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3890..4699
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to kanamycin and geneticin"
                     /protein_id="AIU45940.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      4705..4902
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        5002..5217
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5218..6021
                     /codon_start=1
                     /product="orotidine-5-phosphate (OMP) decarboxylase"
                     /label=orotidine-5-phosphate
                     /note="URA3"
                     /protein_id="AIU45941.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ColE1 pBR322
                     /note="ColE1 pBR322"