RCAS GFP (CT#22) vector (V000422)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V000422	RCAS GFP (CT#22)	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:: RCAS GFP (CT#22)

Antibiotic Resistance:: Ampicillin

Length:: 12448 bp

Type:: Retroviral ; avian expression

Replication origin:: ori

Copy Number:: High Copy

Promoter:: RSV

Cloning Method:: Restriction Enzyme

5' Primer:: RCAS-F

Growth Strain(s):: DH5a

Growth Temperature:: 37℃

RCAS GFP (CT#22) 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

Chen CM, Smith DM, Peters MA, Samson ME, Zitz J, Tabin CJ, Cepko CL. Production and design of more effective avian replication-incompetent retroviral vectors. Dev Biol. 1999 Oct 15;214(2):370-84. doi: 10.1006/dbio.1999.9432. PMID: 10525341.

RCAS GFP (CT#22) vector Sequence

LOCUS       V000422                12448 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000422
VERSION     V000422
KEYWORDS    RCAS GFP (CT#22)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12448)
  AUTHORS   Chen CM, Smith DM, Peters MA, Samson ME, Zitz J, Tabin CJ, Cepko CL
  TITLE     Production and design of more effective avian
            replication-incompetent retroviral vectors.
  JOURNAL   Dev Biol. 1999 Oct 15. 214(2):370-84.
   PUBMED   10525341
REFERENCE   2  (bases 1 to 12448)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 12448)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Biol.
            1999 Oct 15. 214(2):370-84."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(634..650)
                     /label="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)."
     promoter        complement(658..688)
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     primer_bind     complement(731..749)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        817..921
                     /label="AmpR promoter"
     CDS             922..1779
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1953..2541
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     2695..2712
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     misc_feature    complement(2727..2867)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     primer_bind     2953..2975
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2972..3160)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     promoter        4003..4264
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     CDS             6340..6711
                     /gene="gag"
                     /label="Proteinase p15"
                     /note="Proteinase p15 from Avian myeloblastosis associated
                     virus. Accession#: P26315"
     CDS             10372..11109
                     /gene="env"
                     /label="Envelope glycoprotein gp95"
                     /note="Envelope glycoprotein gp95 from Rous sarcoma virus
                     subgroup A (strain Schmidt-Ruppin). Accession#: P0DTM5"
     CDS             11312..12028
                     /label="EGFP"
                     /note="enhanced GFP"
     promoter        12222..12448
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"