LOCUS       Exported               11280 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Lentiviral vector for inducible expression of genes with Gal4DBDVP64
            synNotch Receptors.
ACCESSION   .
VERSION     .
KEYWORDS    pHR_Gal4UAS_IRES_mC_pGK_tBFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11280)
  AUTHORS   Morsut L, Roybal KT, Xiong X, Gordley RM, Coyle SM, Thomson M, Lim 
            WA
  TITLE     Engineering Customized Cell Sensing and Response Behaviors Using 
            Synthetic Notch Receptors.
  JOURNAL   Cell. 2016 Feb 11;164(4):780-91. doi: 10.1016/j.cell.2016.01.012. 
            Epub 2016 Jan 28.
  PUBMED    26830878
REFERENCE   2  (bases 1 to 11280)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.01.012"; journalName: "Cell"; date: 
            "2016-02-11- 11"; volume: "164"; issue: "4"; pages: "780-91"
FEATURES             Location/Qualifiers
     source          1..11280
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             198..908
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(343..361)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     611..630
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1028..1051)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1413..1432
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1501..2202
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    2238..2826
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2291..2311)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2709..2720)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2829..2845)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2830..2846)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2936..3169
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(3194..3212)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3195..3212)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3502..3521)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3735..3754)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3854..3876
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3914..3932)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4000..4104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4105..4965
                     /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(4323..4342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5136..5724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5625..5644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5878..5895
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5965..5985)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5970..6299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6150..6285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6212..6231
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          7433..7498
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7628..7648
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8073..8207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8110..8129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8164..8183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8375..9008
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    9055..9180
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9677..9910
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10095..10139
                     /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    10424..10541
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     10767..10791
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    join(10886..11280,1..179)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(11053..11070)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     join(11280,1..19)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"