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pCS6-Empty vector (V011170)

Price Information

Cat No. Plasmid Name Availability Add to cart
V011170 pCS6-Empty In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

pCS6-Empty is an empty cloning vector for transient mammalian cell expression of a cDNA. It is similar to the pCMV-SPORT6.

Vector Name:
pCS6-Empty
Antibiotic Resistance:
Ampicillin
Length:
4318 bp
Type:
Mammalian Expression Vectors
Replication origin:
ori
Source/Author:
transOMIC
Copy Number:
High copy number
Promoter:
CMV
5' Primer:
M13 fwd
3' Primer:
M13 rev

pCS6-Empty vector Map

Copy Sequence View Map
pCS6-Empty4318 bp600120018002400300036004200SV40 poly(A) signalM13 fwdT7 promoterMCSSP6 promoterM13 revCMV promoterCMV enhanceroriloxPAmpRf1 ori

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

  • Zhang N, Heruth DP, Wu W, Zhang LQ, Nsumu MN, Shortt K, Li K, Jiang X, Wang B, Friesen C, Li DY, Ye SQ. Functional characterization of SLC26A3 c.392C>G (p.P131R) mutation in intestinal barrier function using CRISPR/CAS9-created cell models. Cell Biosci. 2019 May 14;9:40.

pCS6-Empty vector Sequence

LOCUS       pCS6-Empty.        4318 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Empty cloning vector for transient mammalian cell expression of a 
            cDNA. Related to pCMV SPORT6.
ACCESSION   .
VERSION     .
KEYWORDS    pCS6-Empty
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4318)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4318)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(125..259)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     664..680
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        691..709
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    711..774
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(775..793)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(804..820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(946..1149)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(1150..1453)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1913..2501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(2579..2612)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2717..3574)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(3862..4317)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"