载体基本信息 | |||
载体名称 | pWPXLd | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 10455 bp | 载体类型 | Mammalian Expression, Lentiviral |
拷贝数 | High Copy | 克隆方法 | Restriction Enzyme |
5'引物 | See map |
pWPXLd 载体载体图谱
pWPXLd 载体载体序列
LOCUS Exported 10455 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pWPXLd SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10455) TITLE Trono Lab Constitutive Lentiviral Plasmids REFERENCE 2 (bases 1 to 10455) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..10455 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 2..635 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 682..807 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1300..1533 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1718..1762 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 2120..3297 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2351..3288 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3245..3265 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 3345..3462 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" regulatory 3538..3547 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3544..4260 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3544..4260 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3589..3610) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3850..3869) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4197..4218 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4340..4928 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4393..4413) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4811..4822) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5017..5250 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(5275..5293) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5276..5293) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5583..5602) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(5816..5835) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5935..5957 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5995..6013) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6081..6185 /gene="bla" /label=AmpR promoter CDS 6186..7046 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6404..6423) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7217..7805 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7706..7725 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7959..7976 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(8046..8066) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 8051..8380 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8231..8366 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8293..8312 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 9514..9579 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 9709..9729 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 10154..10288 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10191..10210) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10245..10264 /label=EBV-rev /note="SV40 polyA terminator, reverse primer"
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