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pBinGFP2 vector (V012831)

Price Information

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V012831 pBinGFP2 In stock, instant shipping

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

Basic Vector Information

Vector Name:
pBinGFP2
Antibiotic Resistance:
Kanamycin
Length:
14230 bp
Type:
Plant Vectors
Replication origin:
oriV
Host:
Plants
Copy Number:
Low copy number
Promoter:
CaMV 35S (enhanced)
Growth Strain(s):
DH5alpha
Growth Temperature:
37℃

pBinGFP2 vector Map

Copy Sequence View Map
pBinGFP214230 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000oriVKanRtrfALB T-DNA repeatNOS promoterNeoR/KanRNOS terminatorCAP binding sitelac promoterlac operatorM13 revoriVCaMV 35S promoter (enhanced)M13 fwdRB T-DNA repeatTetRoriTtraJ

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.

pBinGFP2 vector Sequence

LOCUS       40924_6532       14230 bp DNA     circular SYN 22-NOV-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14230)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..616
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     CDS             2871..3662
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             3964..5109
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     misc_feature    6708..6732
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        7205..7388
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             7412..8200
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     terminator      8593..8845
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    9394..9415
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9430..9460
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9468..9484
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     9492..9508
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      10000..10624
                     /label=oriV
                     /note="incP origin of replication"
     promoter        complement(10649..11320)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     primer_bind     complement(11439..11455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    11753..11777
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             12242..12889
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     oriT            13573..13682
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             13715..14083
                     /label=traJ
                     /note="oriT-recognizing protein"