RCAS GFP (CT#22) vector (V000422)

Basic Vector Information

      • Vector Name:
      • RCAS GFP (CT#22)
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 12448 bp
      • Type:
      • Retroviral ; avian expression
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • RCAS-F

RCAS GFP (CT#22) vector Vector Map

RCAS GFP (CT#22)12448 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000In lacZ genelac UV5 promoterpBRforEcoAmpR promoterAmpRoriL4440bompGEX 3'RSV promoterEGFPRSV promoter

Plasmid Resuspension 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 ℃.

RCAS GFP (CT#22) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               12448 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Biol. 
            1999 Oct 15. 214(2):370-84."
FEATURES             Location/Qualifiers
     source          1..12448
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(623..645)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    634..650
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             922..1782
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1140..1159)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1953..2541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2442..2461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2695..2712
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2727..2867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2793..2812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2953..2975
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2969..3160)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4003..4264
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     CDS             11312..12031
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             11312..12028
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(11357..11378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(11618..11637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11965..11986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        join(12220..12448,1..33)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"

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