LOCUS       Exported               10074 bp ds-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)
  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."
FEATURES             Location/Qualifiers
     source          1..10074
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(119..137)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(120..137)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1031..1891
                     /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(1249..1268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2062..2650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2551..2570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2804..2821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2891..2911)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2896..3225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3076..3211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4359..4424
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4554..4574
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4999..5133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5036..5055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5090..5109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     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
                     /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"
     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
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             8552..8578
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8669..9379
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(8696..8724)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     9319..9339
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     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"
     LTR             join(9935..10074,1..94)
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"