LOCUS       Exported                6941 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Anti-mouse CD19 CAR with CD3 ITAMs mutated.
ACCESSION   .
VERSION     .
KEYWORDS    MSGV1-1D3-28Z.1-3 mut
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  AUTHORS   Kochenderfer JN, Yu Z, Frasheri D, Restifo NP, Rosenberg SA
  TITLE     Adoptive transfer of syngeneic T cells transduced with a chimeric 
            antigen receptor that recognizes murine CD19 can eradicate lymphoma 
            and normal B cells.
  JOURNAL   Blood. 2010 Nov 11;116(19):3875-86. doi: 
            10.1182/blood-2010-01-265041. Epub 2010 Jul 14.
  PUBMED    20631379
REFERENCE   2  (bases 1 to 6941)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1182/blood-2010-01-265041"; journalName: "Blood"; date: 
            "2010-11-11- 11"; volume: "116"; issue: "19"; pages: "3875-86"
FEATURES             Location/Qualifiers
     source          1..6941
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..517
                     /label=5 LTR
                     /note="5 long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     1357..1379
                     /label=pLXSN 5
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1395..1411
                     /label=pBABE 5
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1749..1765
                     /label=pBMN 5
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1822..1831
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     LTR             3326..3840
                     /label=3 LTR
                     /note="3 long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4012..4028
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4036..4066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4081..4102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4219..4236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4390..4978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4470..4489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5149..6009)
                     /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     5772..5791
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6010..6114)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6182..6200
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6238..6260)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     6360..6379
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6573..6595
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6723..6742
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"