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pCB301 plasmid pXT1 vector (V008700)

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V008700 pCB301 plasmid pXT1 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:
pCB301 plasmid pXT1
Antibiotic Resistance:
Kanamycin
Length:
4623 bp
Type:
Plant transformation vector
Replication origin:
oriV
Source/Author:
Tao X, Yao M, Hu Z, Wei Q, Feng Z.
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pCB301 plasmid pXT1 vector Map

Copy Sequence View Map
pCB301 plasmid pXT14623 bp600120018002400300036004200CaMV 35S promoter (enhanced)HDV-RZNOS terminatorRB T-DNA repeattrfAKanRoriVLB T-DNA repeat

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.

pCB301 plasmid pXT1 vector Sequence

LOCUS       40924_9111        4623 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Plant transformation vector pCB301 plasmid pXT1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4623)
  AUTHORS   Tao X, Yao M, Hu Z, Wei Q, Feng Z.
  TITLE     Construction of Agrobacterium-mediated Cucumber Mosaic Virus 
            Infectious cDNA Clones and 2b Deletion Viral Vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4623)
  AUTHORS   Tao X, Yao M, Hu Z, Wei Q, Feng Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2011) Department of Plant Pathology, Nanjing 
            Agricultural University, No.1 Weigang Street, Nanjing, Jiangsu 
            210095, P.R.China
REFERENCE   3  (bases 1 to 4623)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4623)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-MAY-2011) Department of Plant Pathology, Nanjing Agricultural 
            University, No.1 Weigang Street, Nanjing, Jiangsu 210095, P.R.China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..767
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    768..800
                     /label=multiple cloning site region
                     /note="multiple cloning site region"
     ncRNA           800..887
                     /product="HDV-RZ"
                     /label=ribozyme
                     /note="from hepatitis delta virus"
                     /ncRNA_class="ribozyme"
     terminator      898..1150
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1198..1222
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1377..2522)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(2824..3615)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      complement(3827..4458)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4536..4560
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"