Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000590 | pLB2 CAG P2Gm | 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:
- pLB2 CAG P2Gm
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10656 bp
- Type:
- Mammalian Expression, Mouse Targeting, Lentiviral,
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- Low Copy
- Promoter:
- CAG
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- GFP miR primer (GCTGGAGTTCGTGACCGCC)
pLB2 CAG P2Gm vector 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.
pLB2 CAG P2Gm vector Sequence
LOCUS V000590 10656 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000590
VERSION V000590
KEYWORDS pLB2 CAG P2Gm
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10656)
AUTHORS Stern P, Astrof S, Erkeland SJ, Schustak J, Sharp PA, Hynes RO
TITLE A system for Cre-regulated RNA interference in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2008 Sep 16. 105(37):13895-900.
PUBMED 18779577
REFERENCE 2 (bases 1 to 10656)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 10656)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 2008 Sep 16. 105(37):13895-900."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10656
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(47..66)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 355..372
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 489..510
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 525..555
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 563..579
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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 568..590
/label="M13/pUC Reverse"
/note="In lacZ gene"
primer_bind 587..603
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 587..603
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
promoter 641..868
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 869..1049
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1096..1221
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1720..1953
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2138..2182
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 2331..2372
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2480..2597
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 3119..3422
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3424..3701
/label="chicken beta-actin promoter"
intron 3703..4720
/label="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 4728..4747
/label="pCAG-F"
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 4809..4818
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4815..5411
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label="PuroR"
/note="confers resistance to puromycin"
/translation="MVEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 5311..5331
/label="Puro-F"
/note="Puromycin resistance gene, forward primer"
CDS 5433..5498
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label="F2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 5514..6230
/label="EGFP"
/note="enhanced GFP"
ncRNA 6283..6404
/label="5' miR-30a"
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
ncRNA 6431..6445
/label="miR-30a loop"
/note="loop from the the 71-nt precursor of the human
miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
ncRNA 6473..6599
/label="3' miR-30a"
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
misc_feature 6685..7273
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7276..7292)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7277..7293)
/label="pBluescriptKS"
/note="For pBluescript vector"
polyA_signal 8436..8643
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
LTR 8662..8842
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(8904..9492)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9666..10523)
/label="AmpR"
/note="beta-lactamase"
promoter complement(10524..10628)
/label="AmpR promoter"