pBINPLUS vector (V010906)

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V010906 pBINPLUS 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:
pBINPLUS
Antibiotic Resistance:
Kanamycin
Length:
12396 bp
Type:
Plant Vectors
Replication origin:
oriV
Host:
Plants
Source/Author:
van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema
Selection Marker:
Neomycin/G418(Geneticin)
Copy Number:
High copy number
Promoter:
NOS
5' Primer:
M13 fwd
3' Primer:
M13 rev
Growth Strain(s):
stbl3
Growth Temperature:
37℃

pBINPLUS vector Map

pBINPLUS12396 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000oriVpBR322 ori (truncated)KanRtrfALB T-DNA repeatNOS promoterNeoR/KanRNOS terminatorCAP binding sitelac promoterlac operatorM13 revMCSM13 fwdRB T-DNA repeatoriVoriTtraJ

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.

pBINPLUS vector Sequence

LOCUS       pBINPLUS.       12396 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Improved binary Agrobacterium vector for plant transformation.
ACCESSION   .
VERSION     .
KEYWORDS    pBINPLUS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12396)
  AUTHORS   van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema 
            WJ.
  TITLE     pBINPLUS: an improved plant transformation vector based on pBIN19.
  JOURNAL   Transgenic Res. 1995;4:288-90.
  PUBMED    7655517
REFERENCE   2  (bases 1 to 12396)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 12396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Transgenic 
            Res."; date: "1995"; volume: "4"; pages: "288-90"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     This sequence was reconstructed using the original paper. The 
            orientation of the truncated pBR322 origin is uncertain.
FEATURES             Location/Qualifiers
     source          1..12396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..616
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      1758..2110
                     /direction=RIGHT
                     /label=pBR322 ori (truncated)
                     /note="pBR322 ori (truncated)"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication (truncated)"
     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             7409..8200
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     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 repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     primer_bind     9492..9508
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(9534..9590)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(9605..9621)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9919..9943
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      10000..10624
                     /label=oriV
                     /note="incP origin of replication"
     oriT            11739..11848
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             11881..12249
                     /label=traJ
                     /note="oriT-recognizing protein"