LOCUS       Exported               11423 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Lentiviral vector for expression of anti-Her2 scFv 24D5-7 Gal4VP64 
            synthetic Notch.
ACCESSION   .
VERSION     .
KEYWORDS    pHR_PGK_antiHer24D5-7_synNotch_Gal4VP64
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11423)
  AUTHORS   Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH, 
            Walker WJ, McNally KA, Lim WA
  TITLE     Engineering T Cells with Customized Therapeutic Response Programs 
            Using Synthetic Notch Receptors.
  JOURNAL   Cell. 2016 Oct 6;167(2):419-432.e16. doi: 
            10.1016/j.cell.2016.09.011. Epub 2016 Sep 29.
  PUBMED    27693353
REFERENCE   2  (bases 1 to 11423)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.09.011"; journalName: "Cell"; date: "2016-10-6-
            6"; volume: "167"; issue: "2"; pages: "419-432.e16"
FEATURES             Location/Qualifiers
     source          1..11423
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             64..93
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2287..2436
                     /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"
     primer_bind     complement(2608..2628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3026..3037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3206..3439
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(3464..3482)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3465..3482)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3772..3791)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4005..4024)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4124..4146
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4184..4202)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4270..4374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4375..5235
                     /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(4593..4612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5406..5994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5895..5914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6148..6165
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6235..6255)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6240..6569
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6420..6555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6482..6501
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          7703..7768
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7898..7918
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8343..8477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8380..8399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8434..8453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8645..9278
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    9325..9450
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9947..10180
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10365..10409
                     /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    10694..10811
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        10871..11370
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10906..10929)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     11303..11322
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      join(11418..11423,1..4)
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"