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Pveg vector (V011644)

Price Information

Cat No. Plasmid Name Availability Add to cart
V011644 Pveg 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

Vector Name:
Pveg
Antibiotic Resistance:
Chloramphenicol
Length:
2307 bp
Type:
Synthetic Biology ; Bacillus BioBrick Box
Replication origin:
ori
Copy Number:
High Copy
Promoter:
veg
Cloning Method:
Restriction Enzyme
5' Primer:
na

Pveg vector Map

Copy Sequence View Map
Pveg2307 bp60012001800his operon terminatorL4440orilambda t0 terminatorCmRcat promoterpBRforEcobacterial terminatorBioBrick prefixBioBrick suffix

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.

Pveg vector Sequence

LOCUS       40924_45938        2307 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Very strong constitutive promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2307)
  AUTHORS   Radeck J, Kraft K, Bartels J, Cikovic T, Durr F, Emenegger J, 
            Kelterborn S, Sauer C, Fritz G, Gebhard S, Mascher T
  TITLE     The Bacillus BioBrick Box: generation and evaluation of essential 
            genetic building blocks for standardized work with Bacillus 
            subtilis.
  JOURNAL   J Biol Eng. 2013 Dec 2;7(1):29. doi: 10.1186/1754-1611-7-29.
  PUBMED    24295448
REFERENCE   2  (bases 1 to 2307)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2307)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
            Eng."; date: "2013-12-2"; pages: "
            10.1186/1754-1611-7-29"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..59
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(83..100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(254..842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1025..1119)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1143..1799)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1800..1903)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     primer_bind     1961..1979
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     terminator      complement(1983..2026)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2029..2050
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     misc_feature    2288..2307
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"