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pSELECT-zeo-Lucia vector (V011443) Gene synthesis in pSELECT-zeo-Lucia backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V011443 pSELECT-zeo-Lucia In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pSELECT-zeo-Lucia
Antibiotic Resistance:
Zeocin
Length:
3769 bp
Type:
Luciferase Vectors
Replication origin:
ori
Source/Author:
InvivoGen
Copy Number:
High copy number
Promoter:
EF-1α core
Growth Strain(s):
Top10

pSELECT-zeo-Lucia vector Map

Copy Sequence View Map
pSELECT-zeo-Lucia3769 bp60012001800240030003600poly(A) signalpause siteEF-1-alpha core promoter5' LTR (truncated)Lucia(R)SV40 poly(A) signalbeta-globin poly(A)BleoREM7 promoterCMV promoterCMV enhancerori

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.

pSELECT-zeo-Lucia vector Sequence

LOCUS       pSELECT-zeo-Luci        3769 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian vector for the expression of secreted Lucia(R) luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    pSELECT-zeo-Lucia
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3769)
  AUTHORS   InvivoGen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3769)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    9..57
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    68..159
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     promoter        196..407
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             420..688
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             728..1357
                     /codon_start=1
                     /product="secreted coelenterazine-utilizing luciferase"
                     /label=secreted coelenterazine-utilizing luciferase
                     /note="Lucia(R)"
                     /note="synthetic gene based on luciferases from marine
                     copepods"
                     /translation="MEIKVLFALICIAVAEAKPTEINEDLNIAAVASNFATTDLETDLF
                     TNWETMNVISTDTEQVNTDADRGKLPGKKLPPDVLRELEANARRAGCTRGCLICLSHIK
                     CTPKMKKFIPGRCHTYEGEKESAQGGIGEAIVDIPEIPGFKDKEPLDQFIAQVDLCADC
                     TTGCLKGLANVQCSDLLKKWLPQRCTTFASKIQGRVDKIKGLAGDR"
     polyA_signal    complement(1379..1500)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    complement(1600..1994)
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(2011..2382)
                     /label=BleoR
                     /note="antibiotic-binding protein"
     promoter        complement(2402..2449)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     promoter        complement(2495..2698)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(2699..3002)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3082..3670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"