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pLNCX chick src Y527F vector (V010333)

Price Information

Cat No. Plasmid Name Availability Add to cart
V010333 pLNCX chick src Y527F 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:
pLNCX chick src Y527F
Antibiotic Resistance:
Ampicillin
Length:
8486 bp
Type:
Mammalian Expression, Retroviral
Replication origin:
ori
Selection Marker:
Neomycin (select with G418)
Copy Number:
High Copy
Cloning Method:
Restriction Enzyme
5' Primer:
LNCX primer

pLNCX chick src Y527F vector Map

Copy Sequence View Map
pLNCX chick src Y527F8486 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400CMV enhancerCMV promoterProto-oncogene tyrosine-protein kinase Src3' LTRpGEX 3'bomL4440oriAmpRAmpR promoterpBRforEco5' LTRMMLV Psigag (truncated)NeoR/KanR

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.

pLNCX chick src Y527F vector Sequence

LOCUS       V010333                 8486 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V010333
VERSION     V010333
KEYWORDS    pLNCX chick src Y527F
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8486)
  TITLE     src plasmids
REFERENCE   2  (bases 1 to 8486)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8486)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        169..548
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        549..752
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     749..773
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     CDS             1079..2677
                     /gene="SRC"
                     /label="Proto-oncogene tyrosine-protein kinase Src"
                     /note="Proto-oncogene tyrosine-protein kinase Src from
                     Gallus gallus. Accession#: P00523"
     LTR             2845..3361
                     /label="3' LTR"
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     primer_bind     complement(3465..3487)
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     misc_feature    3573..3713
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3728..3745)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3899..4487)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4661..5518)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5519..5623)
                     /label="AmpR promoter"
     primer_bind     5691..5709
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     LTR             5869..6456
                     /label="5' LTR"
                     /note="long terminal repeat from Moloney murine sarcoma
                     virus"
     misc_feature    6519..6718
                     /label="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             6919..7335
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     CDS             7379..8170
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"