PX853 vector (V012752)

MSDS

Basic Vector Information

N-term SpCas9 piece of inducible split-5 (Cas9(N)-FRB-NES split-5).

- - Vector Name:
  - PX853
- - Antibiotic Resistance:
  - Ampicillin
- - Length:
  - 6295 bp
- - Type:
  - CRISPR/Cas
- - Replication origin:
  - ori
- - Copy Number:
  - High copy number
- - Promoter:
  - CBh

PX853 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 ℃.

PX853 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       PX853.        6295 bp DNA     circular SYN 11-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    PX853.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6295)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        440..725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        727..1004
                     /label=chicken beta-actin promoter
     intron          1005..1233
                     /note="hybrid intron"
                     /note="hybrid between chicken beta-actin (CBA) and minute
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1275..2993
                     /label=Cas9(N)
                     /note="N-terminal portion of Streptococcus pyogenes Cas9 
                     (Zetsche et al., 2015)"
     CDS             3039..3308
                     /label=FRB
                     /note="FKBP-rapamycin binding domain of human FRAP"
     polyA_signal    3342..3549
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3558..3698
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3773..4228
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4510..4614
                     /label=AmpR promoter
     CDS             4615..5472
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5646..6234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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