LOCUS       40924_40622        8872 bp DNA     circular SYN 17-AUG-2021
DEFINITION  Modular 41BB-CD3z CAR backbone, For Transient Expression or 
            Lentiviral Production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8872)
  AUTHORS   Bloemberg D, Nguyen T, MacLean S, Zafer A, Gadoury C, Gurnani K, 
            Chattopadhyay A, Ash J, Lippens J, Harcus D, Page M, Fortin A, Pon 
            RA, Gilbert R, Marcil A, Weeratna RD, McComb S
  TITLE     A High-Throughput Method for Characterizing Novel Chimeric Antigen 
            Receptors in Jurkat Cells.
  JOURNAL   Mol Ther Methods Clin Dev. 2020 Jan 31;16:238-254. doi: 
            10.1016/j.omtm.2020.01.012. eCollection 2020 Mar 13.
  PUBMED    32083149
REFERENCE   2  (bases 1 to 8872)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8872)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther
            Methods Clin Dev."; date: "2020-01-31"; pages: " 
            10.1016/j.omtm.2020.01.012. eCollection 2020 Mar 13"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /codon_start=1
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2128..2245
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2271..2482
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2613..2636
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             4107..4163
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4164..4877
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    4895..5483
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5555..5788
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5866..6000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6027..6162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6183..6201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6211..6227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6369..6824
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6850..6954
                     /label=AmpR promoter
     CDS             6955..7812
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7986..8574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8682..8701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccc gggtttatta cagggacagc
     2101 agagatccac tttggcgccg gctcgagttt taaaagaaaa ggggggattg gggggtacag
     2161 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     2221 acaaattaca aaaattcaaa atttttcgag tggctccggt gcccgtcagt gggcagagcg
     2281 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     2341 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     2401 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     2461 cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg ctcaggctgc
     2521 tcttggctct caacttattc ccttcaattc aagtaacagg agggtcttcg gactacaagg
     2581 atcatgacgg agactataag gatcacgata ttgattacaa agatgacgac gacaaagaca
     2641 tccagatgac acagactaca tcctccctgt ctgcctctct gggagacaga gtcaccatca
     2701 gttgcagggc aagtcaggac atctctaagt atttgaattg gtatcagcag aaaccagatg
     2761 gaactgttaa actcctgatc taccatacat caagattaca ctcaggagtc ccatcaaggt
     2821 tcagtggcag tgggtctgga acagattatt ctctcaccat tagcaacctg gagcaagaag
     2881 atattgccac ttacttttgc caacagggta atacgcttcc gtacacgttc ggagggggga
     2941 ctaagttgga aataacaggc tccacctctg gatccggcaa gcccggatct ggcgagggat
     3001 ccaccaaggg cgaggtgaaa ctgcaggagt caggacctgg cctggtggcg ccctcacaga
     3061 gcctgtccgt cacatgcact gtctcagggg tctcattacc cgactatggt gtaagctgga
     3121 ttcgccagcc tccacgaaag ggtctggagt ggctgggagt aatatggggt agtgaaacca
     3181 catactataa ttcagctctc aaatccagac tgaccatcat caaggacaac tccaagagcc
     3241 aagttttctt aaaaatgaac agtctgcaaa ctgatgacac agccatttac tactgtgcca
     3301 aacattatta ctacggtggt agctatgcta tggactactg gggtcaagga acctcagtca
     3361 ccgtctcctc agcggccgca ggtaccacca caacgcccgc tcctcggcca ccgacgccag
     3421 cgccaactat tgcgagtcag cctctcagtc tgcgacctga ggcttgtcga ccagcagccg
     3481 gaggcgcagt gcacacgagg gggctggact tcgcctgtga tagaagacct ccttctaagc
     3541 ccttttgggt gctggtggtg gttggtggag tcctggcttg ctatagcttg ctagtaacag
     3601 tggcctttat tattttctgg gtgaggaaac ggggcagaaa gaaactcctg tatatattca
     3661 aacaaccatt tatgagacca gtacaaacta ctcaagagga agatggctgt agctgccgat
     3721 ttccagaaga agaagaagga ggatgtgaac tggctagcct gagagtgaag ttcagcagga
     3781 gcgcagacgc ccccgcgtac cagcagggcc agaaccagct ctataacgag ctcaatctag
     3841 gacgaagaga ggagtacgat gttttggaca agagacgtgg ccgggaccct gagatggggg
     3901 gaaagccgca gagaaggaag aaccctcagg aaggcctgta caatgaactg cagaaagata
     3961 agatggcgga ggcctacagt gagattggga tgaaaggcga gcgccggagg ggcaaggggc
     4021 acgatggcct ttaccagggt ctcagtacag ccaccaagga cacctacgac gcccttcaca
     4081 tgcaggccct gccccctcgc gctagcgcca cgaacttctc tctgttaaag caagcaggcg
     4141 acgtggaaga aaaccccggt cccgtgagca agggcgagga gctgttcacc ggggtggtgc
     4201 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4261 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     4321 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     4381 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     4441 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4501 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4561 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4621 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4681 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4741 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4801 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4861 tggacgagct gtacaagtaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     4921 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4981 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5041 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5101 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5161 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5221 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5281 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     5341 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5401 cctgctgccg gctctgcggc ctcttccgcg tctttgcctt cgccctcaga cgagtcggat
     5461 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     5521 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5581 agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     5641 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5701 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5761 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     5821 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     6001 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     6061 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6121 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     6181 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     6241 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     6301 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     6361 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6421 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6481 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6541 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6601 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6661 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6721 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6781 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     6841 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6901 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6961 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7021 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7081 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7141 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     7201 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7261 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7321 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7381 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7441 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7501 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7561 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7621 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7681 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7741 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7801 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8101 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     8161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8281 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtgtcga
     8641 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     8701 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8761 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8821 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
