pCAMBIA2300 vector (V010873)

COA reports Sequencing Results MSDS Download GenBank File(.gb)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V010873	pCAMBIA2300	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pCAMBIA2300

Antibiotic Resistance:: Kanamycin

Length:: 8744 bp

Type:: Plant Vectors

Replication origin:: ori

Host:: Plants

Source/Author:: Cambia

Selection Marker:: Neomycin/G418(Geneticin)

Copy Number:: High copy number

Promoter:: CaMV 35S (enhanced)

5' Primer:: M13 fwd

3' Primer:: M13 rev

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

pCAMBIA2300 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

Sahab S, Taylor N. Studies on Pure Mlb® (Multiple Left Border) Technology and Its Impact on Vector Backbone Integration in Transgenic Cassava. Front Plant Sci. 2022 Feb 4;13:816323. doi: 10.3389/fpls.2022.816323. PMID: 35185986; PMCID: PMC8855067.

Leclercq J, Szabolcs T, Martin F, Montoro P. Development of a new pCAMBIA binary vector using Gateway® technology. Plasmid. 2015 Sep;81:50-4. doi: 10.1016/j.plasmid.2015.07.003. Epub 2015 Jul 23. PMID: 26210260.

Matheka J, Tripathi JN, Merga I, Gebre E, Tripathi L. A simple and rapid protocol for the genetic transformation of Ensete ventricosum. Plant Methods. 2019 Nov 8;15:130. doi: 10.1186/s13007-019-0512-y. PMID: 31719836; PMCID: PMC6839154.

pCAMBIA2300 vector Sequence

LOCUS       Exported                8744 bp DNA     circular SYN 03-SEP-2024
DEFINITION  Agrobacterium binary vector for plant transformation, with 
            kanamycin-resistance genes.
ACCESSION   AF234315
VERSION     .
KEYWORDS    pCAMBIA2300
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8744)
  AUTHORS   Cambia
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8744)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The GenBank record was corrected by inserting a G at position 2281.
FEATURES             Location/Qualifiers
     source          1..8744
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1219..1848
                     /codon_start=1
                     /product="stability protein from Pseudomonas plasmid pVS1"
                     /label=stability protein from Pseudomonas plasmid pVS1
                     /note="pVS1 StaA"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2277..3350
                     /codon_start=1
                     /product="replication protein from Pseudomonas plasmid
                     pVS1"
                     /label=replication protein from Pseudomonas plasmid
                     pVS1
                     /note="pVS1 RepA"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3416..3610
                     /label=pVS1 oriV
                     /note="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3954..4094
                     /label=bom
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4280..4868)
                     /direction=LEFT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4955..5749)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=aphA-3
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6174..6198
                     /label=LB T-DNA repeat
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    6276..6450
                     /label=CaMV poly(A) signal
                     /note="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6507..7304)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=aph(3')-II (or nptII)
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(7367..8044)
                     /note="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     promoter        8271..8301
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    8309..8325
                     /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     8333..8349
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8345..8578
                     /codon_start=1
                     /gene="lacZ (truncated)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ (truncated)
                     /note="lacZ-alpha"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    8359..8415
                     /label=MCS
                     /note="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8419..8435)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8638..8662
                     /label=RB T-DNA repeat
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"

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