pCB301 plasmid pXT1 vector (V008700) Gene synthesis in pCB301 plasmid pXT1 backbone

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V008700	pCB301 plasmid pXT1	In stock, instant shipping

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

Basic Vector Information

RK2 has a low copy-number (around 4–8 copies per chromosome in Escherichia coli) and is stably maintained in diverse Gram negative bacterial species. It is a conjugative plasmid containing oriT in its tra region and an additional trb region encoding a Type IV Secretion system (T4SS) that produces the essential pilus . The other essential backbone functions are a vegetative replication origin (oriV) and a gene encoding a replication initiation protein (trfA), which is regulated by the products of a dual partitioning and central control region (ccr) encoding korA, incC and korB . The broad-host-range and selftransmissibility make RK2 a suitable vector for genetic manipulations and a very good model to study plasmids in bacteria. Examination of RK2 replication intermediates and deletion analysis located the origin required for the initiation of the plasmid vegetative replication, oriV, while trfA, which encodes the essential trans-acting Rep protein, is located upstream from oriV.

Vector Name:: pCB301 plasmid pXT1

Antibiotic Resistance:: Kanamycin

Length:: 4622 bp

Type:: Plant transformation vector

Replication origin:: oriV

Source/Author:: Tao X, Yao M, Hu Z, Wei Q, Feng Z.

Copy Number:: Low copy number

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

pCB301 plasmid pXT1 vector Map

Copy Sequence View Map

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

Wang Q, Zou Q, Dai Z, Hong N, Wang G, Wang L. Four Novel Mycoviruses from the Hypovirulent Botrytis cinerea SZ-2-3y Isolate from Paris polyphylla: Molecular Characterisation and Mitoviral Sequence Transboundary Entry into Plants. Viruses. 2022 Jan 14;14(1):151. doi: 10.3390/v14010151. PMID: 35062353; PMCID: PMC8777694.

pCB301 plasmid pXT1 vector Sequence

LOCUS       pCB301_plasmid_p        4622 bp DNA     circular SYN 20-JUL-2025
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 4622)
  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 4622)
  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 4622)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4622)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4622)
  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"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4622
                     /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
                     MAQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(2824..3615)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      complement(3824..4455)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4535..4559
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"

.cls { stroke-width: 0px; fill: url(#c6); } pCB301 plasmid pXT1 vector (V008700) Gene synthesis in pCB301 plasmid pXT1 backbone