pSR58.6 vector (V001135)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V001135	pSR58.6	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pSR58.6

Antibiotic Resistance:: Chloramphenicol

Length:: 3936 bp

Type:: Bacterial Expression, Synthetic Biology

Replication origin:: ori

Copy Number:: High Copy

pSR58.6 vector Map

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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.

pSR58.6 vector Sequence

LOCUS       pSR58.6.        3936 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses CcaR constitutively and sfGFP under the PcpcG2-172 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pSR58.6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3936)
  AUTHORS   Schmidl SR, Sheth RU, Wu A, Tabor JJ
  TITLE     Refactoring and optimization of light-switchable escherichia coli 
            two-component systems.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):820-31. doi: 10.1021/sb500273n. 
            Epub 2014 Oct 11.
  PUBMED    25250630
REFERENCE   2  (bases 1 to 3936)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3936)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/sb500273n";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "820-31"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      795..866
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      882..909
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1205..1216
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1223..1936
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     terminator      complement(1966..2009)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2050..2077
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2279..2867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2955..3049)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3073..3729)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3730..3832)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     primer_bind     3898..3916
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"