pUC57-sgRNA expression vector vector (V006573)

MSDS

Basic Vector Information

- - Vector Name:
  - pUC57-sgRNA expression vector
- - Antibiotic Resistance:
  - Kanamycin
- - Length:
  - 2792 bp
- - Type:
  - Mammalian Expression, CRISPR
- - Replication origin:
  - ori
- - Copy Number:
  - High Copy
- - Promoter:
  - T7
- - Cloning Method:
  - Restriction Enzyme
- - 5' Primer:
  - pBAC
- - 3' Primer:
  - Lac-promoter

pUC57-sgRNA expression vector vector Vector Map

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 ℃.

pUC57-sgRNA expression vector vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_45193        2792 bp DNA     circular SYN 13-MAY-2021
DEFINITION  For in vitro transcription of sgRNA from the T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2792)
  AUTHORS   Shen B, Zhang W, Zhang J, Zhou J, Wang J, Chen L, Wang L, Hodgkins 
            A, Iyer V, Huang X, Skarnes WC
  TITLE     Efficient genome modification by CRISPR-Cas9 nickase with minimal 
            off-target effects.
  JOURNAL   Nat Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857.
  PUBMED    24584192
REFERENCE   2  (bases 1 to 2792)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2792)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     191..207
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        254..272
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        294..369
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(427..443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(451..467)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(475..505)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(520..541)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(659..676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(830..1418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1592..2404)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2405..2509)
                     /label=AmpR promoter
     primer_bind     2577..2595
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(2633..2655)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2755..2774
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"

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