Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000457 | phage-cmv-cfp-24xms2 | 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:
- phage-cmv-cfp-24xms2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8502 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- WPRE-R (5'-CATAGCGTAAAAGGAGCAACA-3')
phage-cmv-cfp-24xms2 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.
phage-cmv-cfp-24xms2 vector Sequence
LOCUS V000457 8502 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000457
VERSION V000457
KEYWORDS phage-cmv-cfp-24xms2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8502)
AUTHORS Wu B, Chao JA, Singer RH
TITLE Fluorescence fluctuation spectroscopy enables quantitative imaging
of single mRNAs in living cells.
JOURNAL Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19.
PUBMED 22735544
REFERENCE 2 (bases 1 to 8502)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8502)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biophys
J."; date: "2012-06-20"; volume: "102(12)"; pages: "2936-44. Epub
2012 Jun 19"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8502
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2058..2175
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2227..2530
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2531..2734
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2731..2755
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS 2827..3543
/label="ECFP"
/note="enhanced CFP"
misc_RNA 3583..3601
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3622..3640
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3692..3710
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3731..3749
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3801..3819
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3840..3858
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3910..3928
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 3949..3967
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4019..4037
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4058..4076
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4128..4146
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4167..4185
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4237..4255
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4276..4294
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4346..4364
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4457..4475
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4496..4514
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4566..4584
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4605..4623
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4675..4693
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4714..4732
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4784..4802
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 4824..4842
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_feature 4892..5480
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5555..5788
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5907..5925)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5993..6097
/label="AmpR promoter"
CDS 6098..6955
/label="AmpR"
/note="beta-lactamase"
rep_origin 7129..7717
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7871..7888
/label="L4440"
/note="L4440 vector, forward primer"
primer_bind 7980..7999
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
protein_bind 8128..8149
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8164..8194
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 8202..8218
/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 8226..8242
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8292..8311
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"