pC1-REX-YFP vector (V006988)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V006988	pC1-REX-YFP	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:: pC1-REX-YFP

Antibiotic Resistance:: Kanamycin

Length:: 5370 bp

Type:: Mammalian Expression

Replication origin:: ori

Copy Number:: High Copy

Promoter:: CMV

Cloning Method:: Restriction Enzyme

5' Primer:: cggtgggaggtctatataag

3' Primer:: tgggaggttttttaaagcaag

pC1-REX-YFP 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.

pC1-REX-YFP vector Sequence

LOCUS       40924_7881        5370 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Genetically encoded fluorescent indicator for measuring NAD/NADH 
            ratio..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5370)
  AUTHORS   Bilan DS, Matlashov ME, Gorokhovatsky AY, Schultz C, Enikolopov G, 
            Belousov VV
  TITLE     Genetically encoded fluorescent indicator for imaging NAD/NADH ratio
            changes in different cellular compartments.
  JOURNAL   Biochim Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519-9. doi: 
            10.1016/j.bbagen.2013.11.018.
  PUBMED    24286672
REFERENCE   2  (bases 1 to 5370)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5370)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochim 
            Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519-9. doi: 
            10.1016/j.bbagen.2013.11.018."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     1067..1088
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1197..1218)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1458..1477)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2158..2279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2286..2741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2768..2872
                     /label=AmpR promoter
     promoter        2874..3231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3266..4057
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4248..4267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4292..4339
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4668..5256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"