Opie2-PuroR vector (V012836)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012836 Opie2-PuroR In stock, 1 week for quality controls

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

Basic Vector Information

Expresses PuroR and dsRed.

Vector Name:
Opie2-PuroR
Antibiotic Resistance:
Ampicillin
Length:
8646 bp
Type:
Insect Cell Vectors
Replication origin:
ori
Source/Author:
Omar Akbari
Promoter:
IE1
Fusion Tag:
DsRed1
Growth Strain(s):
DH5alpha
Growth Temperature:
37℃

Opie2-PuroR vector Vector Map

Opie2-PuroR8646 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400L4440CAP binding sitelac promoterlac operatorM13 revpiggyBac left (5') inverted repeatSV40 poly(A) signalDsRed1IE1 promoterhr5 enhancerPuroRSV40 poly(A) signalpiggyBac right (3') inverted repeatM13 fwdpRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRori

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.

Opie2-PuroR vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Opie2-PuroR.        8646 bp DNA     circular SYN 15-JAN-2024
DEFINITION  Expresses PuroR and dsRed..
ACCESSION   .
VERSION     .
KEYWORDS    Opie2-PuroR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8646)
  AUTHORS   Kandul NP, Liu J, Hsu AD, Hay BA, Akbari OS
  TITLE     A drug-inducible sex-separation technique for insects.
  JOURNAL   Nat Commun. 2020 Apr 30;11(1):2106. doi: 10.1038/s41467-020-16020-2.
  PUBMED    32355156
REFERENCE   2  (bases 1 to 8646)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun."; date: "2020-04-30"; pages: "
            10.1038/s41467-020-16020-2"
FEATURES             Location/Qualifiers
     source          1..8646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..77
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    194..215
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        230..260
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    268..284
                     /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     292..308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   489..523
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal
                     repeat sequence (ITR)"
     polyA_signal    complement(1212..1333)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1457..2134)
                     /label=DsRed1
                     /note="wild-type DsRed"
     promoter        complement(2145..2736)
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus
                     Autographa californica"
     enhancer        complement(2740..3222)
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     CDS             3785..4381
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     polyA_signal    4402..4536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5572..5634
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal
                     repeat sequence (ITR)"
     primer_bind     complement(6338..6354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6563..6582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     6682..6704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6742..6760)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        6828..6932
                     /label=AmpR promoter
     CDS             6933..7790
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7964..8552
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"