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pG-KJE8 vector (V012941) Gene synthesis in pG-KJE8 backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V012941 pG-KJE8 In stock, instant shipping

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

Basic Vector Information

Vector Name:
pG-KJE8
Antibiotic Resistance:
Chloramphenicol
Length:
11193 bp
Type:
Packaging assistance
Replication origin:
p15A ori
Host:
E. coli
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pG-KJE8 vector Map

Copy Sequence View Map
pG-KJE811193 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000CmRChaperone protein DnaKChaperone protein DnaJProtein GrpErrnB T1 terminatorrrnB T2 terminatorAmpR promoterp15A oriTetRtetR/tetA promotersS loopChaperonin GroELrrnB T1 terminatorcat promoter

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.

pG-KJE8 vector Sequence

LOCUS       Exported               11193 bp DNA     circular SYN 28-NOV-2025
DEFINITION  Exported.
ACCESSION   V012941
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11193)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 11193)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1787..3700
                     /gene="dnaK"
                     /label=Chaperone protein DnaK
                     /note="Chaperone protein DnaK from Escherichia coli O1:K1 /
                     APEC. Accession#: A1A766"
     CDS             3792..4919
                     /gene="dnaJ"
                     /label=Chaperone protein DnaJ
                     /note="Chaperone protein DnaJ from Escherichia coli (strain
                     K12). Accession#: P08622"
     CDS             4939..5529
                     /gene="grpE"
                     /label=Protein GrpE
                     /note="Protein GrpE from Escherichia coli (strain K12). 
                     Accession#: P09372"
     terminator      5835..5921
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6013..6040
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        6060..6150
                     /label=AmpR promoter
     rep_origin      6476..7021
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(7375..7998)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        8014..8069
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             8385..8438
                     /label=S loop
                     /note="GroES chaperone mobile loop that interacts with
                     GroEL"
     CDS             8677..10320
                     /gene="groEL"
                     /label=Chaperonin GroEL
                     /note="Chaperonin GroEL from Escherichia coli O8 (strain
                     IAI1). Accession#: B7M8Q4"
     terminator      10450..10536
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        10872..10974
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             join(10975..11193,1..438)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"