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pIM1773-PRHIII-sfGFP-pNW33N vector (V001611)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001611 pIM1773-PRHIII-sfGFP-pNW33N 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:
pIM1773-PRHIII-sfGFP-pNW33N
Antibiotic Resistance:
Chloramphenicol
Length:
5062 bp
Type:
Bacterial Expression
Replication origin:
ori
Promoter:
PRHIII
Cloning Method:
Restriction Enzyme
3' Primer:
M13-Rev

pIM1773-PRHIII-sfGFP-pNW33N vector Map

Copy Sequence View Map
pIM1773-PRHIII-sfGFP-pNW33N5062 bp6001200180024003000360042004800BioBrick prefixsuperfolder GFPBioBrick suffixM13 revlac operatorlac promoterCAP binding siteChloramphenicol acetyltransferaserepBL4440ori

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.

pIM1773-PRHIII-sfGFP-pNW33N vector Sequence

LOCUS       V001611                 5062 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001611
VERSION     V001611
KEYWORDS    pIM1773-PRHIII-sfGFP-pNW33N
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5062)
  AUTHORS   Blanchard K, Robic S, Matsumura I
  TITLE     Transformable facultative thermophile Geobacillus stearothermophilus
            NUB3621 as a host strain for metabolic engineering.
  JOURNAL   Appl Microbiol Biotechnol. 2014 May 2.
   PUBMED   24788326
REFERENCE   2  (bases 1 to 5062)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5062)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl
            Microbiol Biotechnol. 2014 May 2."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             149..865
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    869..889
                     /label="BioBrick suffix"
                     /note="universal suffix for all parts"
     primer_bind     complement(914..930)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(938..954)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(962..992)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1007..1028)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1161..1808)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(1918..2919)
                     /label="repB"
                     /note="RepB replication protein"
     primer_bind     complement(3440..3457)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3611..4199)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    5062
                     /label="BioBrick prefix"
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"