LOCUS       40924_2079        6941 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Anti-mouse CD19 CAR with CD3 ITAMs mutated.
ACCESSION   .
VERSION     .
KEYWORDS    .
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)
  TITLE     Direct Submission
REFERENCE   3  (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"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     CDS             1020..1427
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="VTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTFNVG
                     WPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKPPPP
                     LPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     misc_feature    1437..1807
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     regulatory      1822..1831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             3326..3840
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     protein_bind    complement(4012..4028)
                     /label=lac operator
                     /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    complement(4081..4102)
                     /label=CAP binding site
                     /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"
     CDS             complement(5152..6009)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6010..6114)
                     /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"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttcgg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttc gggggccgtt tttgtggccc
      781 gacctgagtc ctaaaatccc gatcgtttag gactctttgg tgcacccccc ttagaggagg
      841 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
      901 tgctttcggt ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt
      961 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa tatgggcccg ggctagcctg
     1021 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     1081 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     1141 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     1201 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     1261 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     1321 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     1381 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat
     1441 atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa
     1501 gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg
     1561 aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc
     1621 acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag aacctagaac
     1681 ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc aaagtagacg
     1741 gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg ggtggaccat
     1801 cctctagccc tcgagaagct tgccaccatg ggtgtcccta cccagctcct gggactgctc
     1861 ctgctgtgga tcaccgacgc catctgcgac atccagatga cccagagccc tgccagcctg
     1921 tctaccagcc tgggcgagac agtgaccatc cagtgtcagg ccagcgagga catctactct
     1981 ggcctggctt ggtatcagca gaagcccggc aagagccctc agctgctgat ctacggcgcc
     2041 agcgacctgc aggacggcgt gcctagcaga ttcagcggca gcggctccgg aacccagtac
     2101 agcctgaaga tcaccagcat gcagaccgag gacgagggcg tgtacttctg ccagcaaggc
     2161 ctgacctacc ctagaacctt cggaggaggc accaagctgg aactgaaggg cggaggcgga
     2221 agtggaggcg gaggatctgg cggcggaggc tctgaagtgc agctgcagca gtctggcgct
     2281 gaactggtcc ggcctggcac tagcgtgaag ctgtcctgca aggtgtccgg cgacaccatc
     2341 accttctact acatgcactt cgtgaagcag aggccaggac agggcctgga atggatcggc
     2401 agaatcgacc ctgaggacga gagcaccaag tacagcgaga agttcaagaa caaggccacc
     2461 ctgaccgccg acaccagcag caacaccgcc tacctgaagc tgtctagcct gacctccgag
     2521 gacaccgcca cctacttttg catctacggc ggctactact tcgactactg gggccagggc
     2581 gtgatggtca ccgtgtccag catcgagttc atgtaccccc ctccctacct ggacaacgag
     2641 agaagcaacg gcaccatcat ccacatcaaa gaaaagcacc tgtgccacac ccagagcagc
     2701 cccaagctgt tctgggccct ggtggtggtg gccggcgtgc tgttctgtta cggcctgctg
     2761 gtcacagtgg ccctgtgcgt gatctggacc aacagcagaa gaaacagagg cggccagagc
     2821 gactacatga acatgacccc cagaaggcca ggcctgacca gaaagcccta ccagccctac
     2881 gcccctgcca gagacttcgc cgcctacaga cccagagcca agttcagcag atccgccgag
     2941 acagccgcca acctgcagga tcccaaccag ctgttcaacg agctgaacct gggcagacgg
     3001 gaggaattcg acgtgctgga aaagaagaga gccagggacc ccgagatggg cggcaagcag
     3061 cagagaagaa gaaaccctca ggaaggcgtc tacaacgccc tgcagaaaga caagatggcc
     3121 gaggcctaca gcgagatcgg caccaagggc gagagaagaa ggggcaaggg ccacgatggc
     3181 ctgttccagg gcctgtccac cgccaccaag gacaccttcg acgccctgca catgcagacc
     3241 ctggccccca gatgagtcga cggtaccgcg ggcccgggat ccgataaaat aaaagatttt
     3301 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     3361 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     3421 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     3481 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     3541 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     3601 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3661 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     3721 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     3781 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     3841 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     3901 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatccat
     3961 cgatggagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc
     4021 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4081 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4141 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4201 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4801 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6001 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6121 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6181 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6241 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6301 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6361 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6421 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6481 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6541 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     6601 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6661 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6721 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6781 acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc
     6841 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     6901 ataaaataaa agattttatt tagtctccag aaaaaggggg g
//
