pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS 载体 (V005190#)

载体名称:
pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS
载体抗性:
Ampicillin
载体长度:
10074 bp
载体类型:
Mammalian Expression, Lentiviral
复制子:
ori
拷贝数:
High Copy
启动子:
SV40

pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS 载体载体图谱

pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS10074 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000SP6 promoterpBRrevBampGEX 3'pBRforEcoAmpR promoterAmpRoriL4440SV40 promotersmall t intronSV40 NLSSV40 poly(A) signal3' LTRHIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSSV40 promoterKozak sequenceHAsuperfolder GFPVP64GB1Rex NLS

pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS 载体载体序列

LOCUS       pHRdSV40-scFv-GC       10074 bp DNA     circular SYN 13-MAY-2021
DEFINITION  The plasmid encodes a antibody that binds to the GCN4 peptide from 
            the SunTag system, and is fused to a transcriptional activation 
            domain VP64.
ACCESSION   .
VERSION     .
KEYWORDS    pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10074)
  AUTHORS   Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, Vale RD
  TITLE     A Protein-Tagging System for Signal Amplification in Gene Expression
            and Fluorescence Imaging.
  JOURNAL   Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 
            10.1016/j.cell.2014.09.039.
  PUBMED    25307933
REFERENCE   2  (bases 1 to 10074)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10074)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2014 
            Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(119..137)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(427..446)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     primer_bind     780..802
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(840..858)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        926..1030
                     /label=AmpR promoter
     CDS             1031..1888
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2062..2650
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2804..2821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2896..3225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     intron          4359..4424
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4554..4574
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    4999..5133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             5302..5935
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5982..6107
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6599..6832
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7017..7061
                     /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             7210..7251
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    7346..7463
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        7526..7736
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7595..7730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      7805..7814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8552..8578
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             8669..9379
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     CDS             9413..9562
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             9593..9754
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             9782..9832
                     /codon_start=1
                     /product="nuclear localization signal from the human T-cell
                     leukemia virus type 1 (HTLV-1) Rex protein"
                     /label=Rex NLS
                     /translation="PKTRRRPRRSQRKRPPT"

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