Cart 2

pCMV-Script vector (V011243)

Price Information

Cat No. Plasmid Name Availability Add to cart
V011243 pCMV-Script 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:
pCMV-Script
Antibiotic Resistance:
Kanamycin
Length:
4278 bp
Type:
Mammalian Expression Vectors
Replication origin:
ori
Source/Author:
Agilent Technologies
Selection Marker:
Neomycin/G418(Geneticin)
Copy Number:
High copy number

pCMV-Script vector Map

Copy Sequence View Map
pCMV-Script4278 bp600120018002400300036004200CMV enhancerCMV promoterT3 promoterMCST7 promoterSV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRHSV TK poly(A) signalori

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.

pCMV-Script vector Sequence

LOCUS       40924_11755        4278 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCMV-Script, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4278)
  AUTHORS   Hosfield T, Padgett K, Sanchez T, Lu Q.
  TITLE     Mammalian expression vector for efficient cloning of PCR fragments
  JOURNAL   Strategies 10, 68-69 (1997)
REFERENCE   2  (bases 1 to 4278)
  AUTHORS   Hosfield T, Padgett K, Sanchez T, Lu Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-OCT-1997) Marketing Analysis, Stratagene, 11011 North 
            Torrey Pines Rd, La Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 4278)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4278)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Strategies 
            10, 68-69 (1997)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-OCT-1997) Marketing Analysis, Stratagene, 11011 North Torrey 
            Pines Rd, La Jolla, CA 92037, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    651..758
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(780..798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1072..1193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1200..1655)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1682..1784
                     /label=AmpR promoter
     promoter        1788..2145
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2180..2971
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3206..3253
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3582..4170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"