pCAMBIA0380 vector (V008771)

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

Basic Vector Information

Vector Name:
pCAMBIA0380
Antibiotic Resistance:
Kanamycin
Length:
6812 bp
Type:
Binary vector
Replication origin:
ori
Source/Author:
Hajdukiewicz P, Svab Z, Maliga P.

pCAMBIA0380 vector Map

pCAMBIA03806812 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006xHisNOS terminatorRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanRLB T-DNA repeatpUC8 MCS; polylinker

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.

pCAMBIA0380 vector Sequence

LOCUS       40924_8676        6812 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCAMBIA-0380, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  AUTHORS   Hajdukiewicz P, Svab Z, Maliga P.
  TITLE     The small, versatile pPZP family of Agrobacterium binary vectors for
            plant transformation
  JOURNAL   Plant Mol. Biol. 25 (6), 989-994 (1994)
  PUBMED    7919218
REFERENCE   2  (bases 1 to 6812)
  AUTHORS   Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
            KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet 
            I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A, 
            Jefferson RA.
  TITLE     A comprehensive set of modular vectors for advanced manipulations 
            and efficient transformation of plants
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6812)
  AUTHORS   Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
            KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet 
            I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A, 
            Jefferson RA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / 
            GPO Box 3200, Canberra, ACT 2601, Australia
REFERENCE   4  (bases 1 to 6812)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6812)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol. 
            Biol."; date: "1994"; volume: "25"; issue: "6"; pages: "989-994"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / GPO Box 
            3200, Canberra, ACT 2601, Australia"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..23
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      58..310
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    332..356
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1656..2282
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             2719..3783
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      3852..4046
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4390..4530
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4716..5304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5394..6185)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    6610..6634
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(6737..6772)
                     /note="pUC8 MCS; polylinker"