LOCUS       Exported               11496 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Lentivector to express STING and mTagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    pTRIP-SFFV-mtagBFP-2A STING
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11496)
  AUTHORS   Cerboni S, Jeremiah N, Gentili M, Gehrmann U, Conrad C, Stolzenberg 
            MC, Picard C, Neven B, Fischer A, Amigorena S, Rieux-Laucat F, Manel
            N
  TITLE     Intrinsic antiproliferative activity of the innate sensor STING in T
            lymphocytes.
  JOURNAL   J Exp Med. 2017 Jun 5;214(6):1769-1785. doi: 10.1084/jem.20161674. 
            Epub 2017 May 8.
  PUBMED    28484079
REFERENCE   2  (bases 1 to 11496)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1084/jem.20161674"; journalName: "J Exp Med"; date: "2017-06-5- 
            5"; volume: "214"; issue: "6"; pages: "1769-1785"
FEATURES             Location/Qualifiers
     source          1..11496
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          43..108
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             238..258
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    683..817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(720..739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     774..793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             985..1618
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    1665..1790
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2283..2516
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2701..2745
                     /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    3038..3155
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3295..3702
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3721..4419
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             4429..4485
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     LTR             5900..6080
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7608..7627)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7841..7860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7960..7982
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8020..8038)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8106..8210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8211..9071
                     /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(8429..8448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9242..9830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9731..9750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9984..10001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10071..10091)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10076..10405
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10256..10391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10318..10337
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"