pBBR1MCS5-Tac-EGFP vector (V012799)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V012799	pBBR1MCS5-Tac-EGFP	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

pBBR1MCS vectors are small in size, contain unique cloning sites within the lacZα gene, and are mobilizable and compatible with various plasmid incompatibility groups.

Vector Name:: pBBR1MCS5-Tac-EGFP

Antibiotic Resistance:: Gentamicin

Length:: 5539 bp

Type:: Expression plasmid

Replication origin:: pBBR1 oriV

Copy Number:: Low copy number

Promoter:: tac

Cloning Method:: Enzyme Cut

3' Primer:: M13 rev

Fusion Tag:: EGFP

Growth Strain(s):: JM108

Growth Temperature:: 37℃

pBBR1MCS5-Tac-EGFP vector Vector Map

View Map in NovoBuilder

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

Zhang BN, Qi B, Chu WK, Song F, Li S, Dong Q, Shao Z, Zhang B, Du X, Ma X, Jhanji V, Zhou Q. Norepinephrine as the Intrinsic Contributor to Contact Lens-Induced Pseudomonas aeruginosa Keratitis. Invest Ophthalmol Vis Sci. 2023 May 1;64(5):26. doi: 10.1167/iovs.64.5.26. PMID: 37234000; PMCID: PMC10226609.

pBBR1MCS5-Tac-EGFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_6073        5539 bp DNA     circular SYN 18-APR-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5539)
  AUTHORS   NOVOPRO
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3239..3255
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3257..3285
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             3315..4031
                     /label=EGFP
                     /note="enhanced GFP"
     primer_bind     complement(4060..4076)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4106..4124)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4145..4161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4169..4185)
                     /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(4193..4223)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4238..4259)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4529..4557
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4746..5276
                     /label=GmR
                     /note="gentamycin acetyltransferase"