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pTf16 vector (V012943)

Price Information

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V012943 pTf16 In stock, instant shipping

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

Basic Vector Information

The pTf16 vector plasmid is a chaperone plasmid. Primarily for facilitating the production and folding of recombinant proteins in bacterial systems, particularly Escherichia coli (E. coli). It has been shown that the co-expression of ScFvs and the chaperone plasmids PG-tf2, pTf16, pGro7, pKJE7, and pG-KJE8 leads to a noticeable increase in the expression of soluble proteins.

Vector Name:
pTf16
Antibiotic Resistance:
Chloramphenicol
Length:
5014 bp
Type:
Packaging assistance
Replication origin:
p15A ori
Host:
E. coli
Growth Strain(s):
DH10B
Growth Temperature:
37℃

pTf16 vector Map

Copy Sequence View Map
pTf165014 bp6001200180024003000360042004800CmRAraCTrigger Factorp15A oricat 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.

References

  • Nazari A, Farajnia S, Zahri S, Bagherlou N, Tanoumand A, Rahbarnia L. Cytoplasmic Chaperones Enhance Soluble Expression of Anti-EGFR huscFv in Escherichia coli. Iran J Biotechnol. 2020 Apr 1;18(2):e2314. doi: 10.30498/IJB.2020.138200.2314. PMID: 33542937; PMCID: PMC7856399.

pTf16 vector Sequence

LOCUS       Exported                5014 bp DNA     circular SYN 10-OCT-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5014)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(574..1419)
                     /codon_start=1
                     /label=AraC
                     /note="arabinose operon transcriptional regulator AraC"
                     /translation="MAETQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGEGVINNNGEQFVCRPGDILLFPPGEIHHYGRHPDASEWYHQWVYFRPRAYW
                     QEWLTWPTIFAQTGFFRPDEARQPHFSELFGQIISAGQGEGRYSELLAINLLEQLLLRR
                     MAVINESLHPPMDSRVRDACQYISDHLADSHFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     "
     CDS             1891..3186
                     /codon_start=1
                     /label=Trigger Factor
                     /note="ribosome-associated chaperone from E. coli (Hoffmann
                     et al., 2010)"
                     /translation="MQVSVETTQGLGRRVTITIAADSIETAVKSELVNVAKKVRIDGFR
                     KGKVPMNIVAQRYGASVRQDVLGDLMSRNFIDAIIKEKINPAGAPTYVPGEYKLGEDFT
                     YSVEFEVYPEVELQGLEAIEVEKPIVEVTDADVDGMLDTLRKQQATWKEKDGAVEAEDR
                     VTIDFTGSVDGEEFEGGKASDFVLAMGQGRMIPGFEDGIKGHKAGEEFTIDVTFPEEYH
                     AENLKGKAAKFAINLKKVEERELPELTAEFIKRFGVEDGSVEGLRAEVRKNMERELKSA
                     IRNRVKSQAIEGLVKANDIDVPAALIDSEIDVLRRQAAQRFGGNEKQALELPRELFEEQ
                     AKRRVVVGLLLGEVIRTNELKADEERVKGLIEEMASAYEDPKEVIEFYSKNKELMDNMR
                     NVALEEQAVEAVLAKAKVTEKETTFNELMNQQA"
     rep_origin      3622..4167
                     /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."
     promoter        4693..4795
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             join(4796..5014,1..438)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"