pMRI-32 vector (V004469)

Basic Vector Information

Vector Name:
pMRI-32
Antibiotic Resistance:
Kanamycin
Length:
4902 bp
Type:
Cloning vector
Replication origin:
ori
Host:
Yeast
Source/Author:
Xie W, Yu H.
Promoter:
GAL1,10

pMRI-32 vector Vector Map

pMRI-324902 bp6001200180024003000360042004800loxPTy4 homologous armADH1 terminatorFLAGGAL1,10 promoterT7 promoterMycCYC1 terminatorT7 promoterLoxp sitekanMXori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pMRI-32 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_32270        4902 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-32, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4902)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2014) Department of Chemical and Biological Engineering, 
            Zhejiang University, Institute of Bioengineering, 34 TianMuShan 
            Road, Hangzhou, Zhejiang 310028, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..55
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    113..1227
                     /label=Ty4 homologous arm
                     /note="Ty4 homologous arm"
     terminator      complement(1262..1427)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1575..1598)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        1645..2309
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        2320..2338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2356..2385
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      2415..2604
                     /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)"
     rep_origin      complement(4226..4814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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