LOCUS       sequence-377864-        4586 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pHR-OFF VIPER CAR (anti-CD19)-candidate design(d+iii).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4586)
  AUTHORS   Li HS, Wong NM, Tague E, Ngo JT, Khalil AS, Wong WW
  TITLE     High-performance multiplex drug-gated CAR circuits.
  JOURNAL   Cancer Cell. 2022 Aug 26. pii: S1535-6108(22)00372-5. doi: 
            10.1016/j.ccell.2022.08.008.
  PUBMED    36084652
REFERENCE   2  (bases 1 to 4586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer 
            Cell. 2022 Aug 26. pii: S1535-6108(22)00372-5. doi: 
            10.1016/j.ccell.2022.08.008."
COMMENT     Sequence Label: pHR-OFF VIPER CAR (anti-CD19)-candidate
            design...
FEATURES             Location/Qualifiers
     source          1..4586
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        6..1184
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          237..1175
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1132..1152
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1221..1230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2016..2057
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2688..2744
                     /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             3876..4586
                     /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(4021..4039)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4289..4308
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
       61 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
      121 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      181 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
      241 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
      301 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
      361 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
      421 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
      481 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
      541 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
      601 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
      661 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
      721 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
      781 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
      841 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
      901 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
      961 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     1021 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     1081 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     1141 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggggat ctggagctct
     1201 cgagaattct cacgcgtgcc gccaccatgg ccttaccagt gaccgccttg ctcctgccgc
     1261 tggccttgct gctccacgcc gccaggccgg acatccagat gacacagact acatcctccc
     1321 tgtctgcctc tctgggagac agagtcacca tcagttgcag ggcaagtcag gacattagta
     1381 aatatttaaa ttggtatcag cagaaaccag atggaactgt taaactcctg atctaccata
     1441 catcaagatt acactcagga gtcccatcaa ggttcagtgg cagtgggtct ggaacagatt
     1501 attctctcac cattagcaac ctggagcaag aagatattgc cacttacttt tgccaacagg
     1561 gtaatacgct tccgtacacg ttcggaggcg gcaccaagct ggagatcaca ggtggcggtg
     1621 gctcgggcgg tggtgggtcg ggtggcggcg gatctgaggt gaaactgcag gagtcaggac
     1681 ctggcctggt ggcgccctca cagagcctgt ccgtcacatg cactgtctca ggggtctcat
     1741 tacccgacta tggtgtaagc tggattcgcc agcctccacg aaagggtctg gagtggctgg
     1801 gagtaatatg gggtagtgaa accacatact ataattcagc tctcaaatcc agactgacca
     1861 tcatcaagga caactccaag agccaagttt tcttaaaaat gaacagtctg caaactgatg
     1921 acacagccat ttactactgt gccaaacatt attactacgg tggtagctat gctatggact
     1981 actggggcca aggaacctca gtcaccgtct cctcagggaa gcctatcccg aaccctctgt
     2041 tgggtctcga tagtaccaat ggggccgcaa ccacgacgcc agcgccgcga ccaccaacac
     2101 cggcgcccac catcgcgttg cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg
     2161 cggggggcgc agtgcacacg agggggctgg acttcgcctg tgatttttgg gtgctggtgg
     2221 tggttggtgg agtcctggct tgctatagct tgctagtaac agtggccttt attattttct
     2281 gggtgaggag taagaggagc aggctcctgc acagtgacta catgaacatg actccccgcc
     2341 gccccgggcc cacccgcaag cattaccagc cctatgcccc accacgcgac ttcgcagcct
     2401 atcgctccaa acggggcaga aagaaactcc tgtatatatt caaacaacca tttatgagac
     2461 cagtacaaac tactcaagag gaagatggct gtagctgccg atttccagaa gaagaagaag
     2521 gaggatgtga actggggggt accgaattca gcgggtcagg cggatctgga agtggtggtg
     2581 gagaacttga tgaattggta tacttactag atgggccagg ttatgaccct atacattgcg
     2641 atgtagtgac aaggggcggc agccaccttt tcaattttgg aagcggagct actaacttca
     2701 gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctatgatt catcttggtc
     2761 acatcctctt cttgctgctg ttgccagttg ctgccgcaca aacaacccca ggtgagaggt
     2821 caagtcttcc cgccttctat ccgggcacaa gcgggagttg tagtgggtgt ggatctttgt
     2881 cccttcctat atacatctgg gctcctctgg ctggcacttg cggagtgctt ctgctgagtc
     2941 tggttattac cggttctgtt gttattgttg gtagaattat tttatctggt agtggtagta
     3001 tcacggccta ctcccaacag acgcggggcc tacttggttg catcatcact agcctcacag
     3061 gccgggacaa gaaccaggtc gaaggggagg ttcaagtggt ttctaccgca acacaatctt
     3121 tcctggcgac ctgcgtcaac ggcgtgtgct ggactgtcta ccatggcgct ggctcgaaga
     3181 ccctagccgg tccaaaaggt ccaatcaccc aaatgtacac caatgtagac caggacctcg
     3241 tcggctggca ggcgcctcca ggggcgcgct ccttgacacc atgcacctgt ggcagctcgg
     3301 acctttactt ggtcacgaga catgctgatg tcattccggt gcgccggcga ggcgacagca
     3361 ggggaagtct actctccccc aggcccgtct cctacctgaa aggctccgca ggtggtccat
     3421 tgctttgccc ttcggggcac gctgtgggca tcttccgggc tgctgtgtgc acccgggggg
     3481 tcgcgaaggc ggtggacttc gtgcccgttg agtctatgga aactaccatg cggtctagag
     3541 tgaagttcag caggagcgca gacgcccccg cgtacaagca gggccagaac cagctctata
     3601 acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga cgtggccggg
     3661 accctgagat ggggggaaag ccgagaagga agaaccctca ggaaggcctg tacaatgaac
     3721 tgcagaaaga taagatggcg gaggcctaca gtgagattgg gatgaaaggc gagcgccgga
     3781 ggggcaaggg gcacgatggc ctttaccagg gtctcagtac agccaccaag gacacctacg
     3841 acgcccttca catgcaggcc ctgccccctc gcggcatggt gagcaagggc gaggaggata
     3901 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     3961 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     4021 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     4081 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     4141 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     4201 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     4261 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     4321 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     4381 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     4441 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     4501 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     4561 gcggcatgga cgagctgtac aagtaa
//
