Basic Vector Information
- Vector Name:
- pLV3-8×GTIIC-mCherry-Fluc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8712 bp
- Type:
- Luciferase reporter
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- hPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-8×GTIIC-mCherry-Fluc vector Vector Map
Plasmid Resuspension 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 ℃.
pLV3-8×GTIIC-mCherry-Fluc vector Sequence
LOCUS 62056_14955 8712 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8712) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..8712 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 7..507 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" CDS 522..2171 /codon_start=1 /label=luciferase /note="firefly luciferase" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL" LTR 2245..2478 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2550..2684 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 2711..2846 /label=SV40 ori /note="SV40 origin of replication" promoter complement(2867..2885) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2895..2911) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 3053..3508 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3534..3638 /label=AmpR promoter CDS 3639..4496 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4670..5258 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5546..5567 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 5582..5612 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5620..5636 /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 5644..5660 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 5681..5699 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5727..5953 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" misc_feature 6181..6306 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6799..7032 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7217..7261 /codon_start=1 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /translation="KNEQELLELDKWASL" primer_bind 7632..7648 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS 7794..8501 /codon_start=1 /label=mCherry /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" misc_feature 8543..8660 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 8709..8712 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter"
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