pCAMBIA1300-35S-VAC-mCherry vector (V013370)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V013370	pCAMBIA1300-35S-VAC-mCherry	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pCAMBIA1300-35S-VAC-mCherry

Antibiotic Resistance:: Kanamycin

Length:: 11918 bp

Type:: Subcellular localization

Replication origin:: ori

Host:: Plants

Selection Marker:: Hyg

Promoter:: CaMV35S(long)

Growth Strain(s):: DH5a

Growth Temperature:: 37℃

pCAMBIA1300-35S-VAC-mCherry vector Map

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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.

pCAMBIA1300-35S-VAC-mCherry vector Sequence

LOCUS       V013370                11918 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013370
VERSION     V013370
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11918)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..17)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    220..244
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1544..2170
                     /label="pVS1 StaA"
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             2607..3671
                     /label="pVS1 RepA"
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      3740..3934
                     /label="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4278..4418
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4604..5192)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5282..6073)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    6498..6522
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(6600..6774)
                     /label="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6817..7839)
                     /label="HygR"
                     /note="aminoglycoside phosphotransferase from E. coli"
     promoter        complement(7907..8584)
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    8775..8796
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8811..8841
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        9398..9743
                     /label="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             9758..10510
                     /gene="TIP1-1"
                     /label="Aquaporin TIP1-1"
                     /note="Aquaporin TIP1-1 from Arabidopsis thaliana.
                     Accession#: P25818"
     CDS             10540..11247
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     terminator      11270..11911
                     /label="E9 terminator"
                     /note="terminator and polyadenylation signal from the pea
                     rbcS-E9 gene"