LOCUS       Exported               10765 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Lentiviral vector - tBFP reporter for Gal4DBDVP64 synNotch receptors
            with a constitutive mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    pHR_Gal4UAS_tBFP_PGK_mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10765)
  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 10765)
  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..10765
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          455..520
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             650..670
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1095..1229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1132..1151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1186..1205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1397..2030
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    2077..2202
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2699..2932
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3117..3161
                     /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    3446..3563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     3789..3813
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3894..4595
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        4718..5217
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4753..4776)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5150..5169
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5238..5948
                     /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(5383..5401)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5651..5670
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6025..6613
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6078..6098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6496..6507)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6616..6632)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6617..6633)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6723..6956
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(6981..6999)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6982..6999)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7289..7308)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7522..7541)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7641..7663
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7701..7719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7787..7891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7892..8752
                     /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(8110..8129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8923..9511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9412..9431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9665..9682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(9752..9772)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9757..10086
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9937..10072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9999..10018
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"