Basic Vector Information
- Vector Name:
- pLV3-SV40-hRluc-PGK-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8522 bp
- Type:
- Luciferase reporter
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-SV40-hRluc-PGK-Puro 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-SV40-hRluc-PGK-Puro vector Sequence
LOCUS 62056_15620 8522 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 8522) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..8522 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 449..574 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1067..1300 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1485..1529 /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" misc_feature 1713..1830 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 1879..2236 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2287..3219 /codon_start=1 /label=hRluc /note="Renilla luciferase" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" promoter 3234..3733 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 3754..4350 /codon_start=1 /label=PuroR /note="puromycin N-acetyltransferase" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 4369..4957 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 5029..5262 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5340..5474 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(5657..5675) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5685..5701) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 5843..6298 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 6324..6428 /label=AmpR promoter CDS 6429..7286 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 7460..8048 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 8336..8357 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 8372..8402 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8410..8426 /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 8434..8450 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 8471..8489 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 8517..8522 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter"
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