LOCUS       Exported               11707 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  [Edit] Lentiviral vector for Gal4 inducible Blinatumomab and 
            constitutive mCherry expression.
ACCESSION   .
VERSION     .
KEYWORDS    pHR_Gal4UAS_Blinatumomab_PGK_mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11707)
  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 11707)
  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..11707
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     166..190
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      271..280
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             343..372
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1867..1884
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1885..1911
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        2037..2536
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2072..2095)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2469..2488
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2557..3267
                     /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(2702..2720)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2970..2989
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3344..3932
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3397..3417)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3815..3826)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3935..3951)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3936..3952)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4042..4275
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(4300..4318)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4301..4318)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4608..4627)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4841..4860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4960..4982
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5020..5038)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5106..5210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5211..6071
                     /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(5429..5448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6242..6830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6731..6750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6984..7001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(7071..7091)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7076..7405
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7256..7391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7318..7337
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          8539..8604
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8734..8754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9179..9313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9216..9235)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9270..9289
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9481..10114
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    10161..10286
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10783..11016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11201..11245
                     /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    11530..11647
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"