pCMV2-SEP-GluA1 (M1) vector (V006807)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V006807	pCMV2-SEP-GluA1 (M1)	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:: pCMV2-SEP-GluA1 (M1)

Antibiotic Resistance:: Ampicillin

Length:: 8314 bp

Type:: Mammalian Expression

Replication origin:: ori

Promoter:: CMV

5' Primer:: CMV-F

3' Primer:: hGH-pA-R

pCMV2-SEP-GluA1 (M1) 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.

pCMV2-SEP-GluA1 (M1) vector Sequence

LOCUS       V006807                 8314 bp    DNA     circular SYN 04-NOV-2021
DEFINITION  Exported.
ACCESSION   V006807
VERSION     V006807
KEYWORDS    pCMV2-SEP-GluA1 (M1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8314)
  TITLE     Jeremy Henley Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8314)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8314)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(293..915)
                     /label="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     CDS             complement(933..3599)
                     /gene="Gria1"
                     /label="Glutamate receptor 1"
                     /note="Glutamate receptor 1 from Rattus norvegicus.
                     Accession#: P19490"
     CDS             complement(3603..4313)
                     /label="superecliptic pHluorin"
                     /note="pH-sensitive mutant of green fluorescent protein (Ng
                     et al., 2002)"
     promoter        complement(4496..4699)
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(4700..5079)
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5240..5256)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      complement(5397..5852)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5879..5983
                     /label="AmpR promoter"
     CDS             5984..6841
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7015..7603
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    7891..7912
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7927..7957
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7965..7981
                     /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)."
     primer_bind     7989..8005
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        8019..8037
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          complement(8114..8210)
                     /label="SV40 intron"
                     /note="modified SV40 intron with splice donor and acceptor
                     sites"
     promoter        complement(join(8249..8314,1..264))
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"