LOCUS       sequence-367846-        5775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BCMA-CAR DNA production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5775)
  AUTHORS   Shy BR, Vykunta VS, Ha A, Talbot A, Roth TL, Nguyen DN, Pfeifer WG, 
            Chen YY, Blaeschke F, Shifrut E, Vedova S, Mamedov MR, Chung JJ, Li 
            H, Yu R, Wu D, Wolf J, Martin TG, Castro CE, Ye L, Esensten JH, 
            Eyquem J, Marson A
  TITLE     High-yield genome engineering in primary cells using a hybrid ssDNA 
            repair template and small-molecule cocktails.
  JOURNAL   Nat Biotechnol. 2022 Aug 25:10.1038/s41587-022-01418-8. doi: 
            10.1038/s41587-022-01418-8.
  PUBMED    36008610
REFERENCE   2  (bases 1 to 5775)
  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: "Nat 
            Biotechnol. 2022 Aug 25:10.1038/s41587-022-01418-8. doi: 
            10.1038/s41587-022-01418-8."
COMMENT     Sequence Label: pIG-641 JE-TRAC-BCMA(BB)28z-P2A
FEATURES             Location/Qualifiers
     source          1..5775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(65..653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(145..164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(824..1615,1616..1684))
                     /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     1447..1466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1685..1789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1857..1875
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1913..1935)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2035..2054
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2071..2526)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2208..2229)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2420..2439
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2601..2741
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2612..2741
                     /label=AAV2 ITR
     CDS             3408..3464
                     /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             4266..4295
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4965..5021
                     /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"
     repeat_region   join(5649..5775,1..3)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
       61 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1801 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     1861 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     1921 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     1981 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2041 cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat
     2101 tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa
     2161 tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca
     2221 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2281 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     2341 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2401 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa
     2461 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2521 gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2581 aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     2641 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     2701 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcta
     2761 gcgatgtaag gagctgctgt gacttgctca aggccttata tcgagtaaac ggtagtgctg
     2821 gggcttagac gcaggtgttc tgatttatag ttcaaaacct ctatcaatga gagagcaatc
     2881 tcctggtaat gtgatagatt tcccaactta atgccaacat accataaacc tcccattctg
     2941 ctaatgccca gcctaagttg gggagaccac tccagattcc aagatgtaca gtttgctttg
     3001 ctgggccttt ttcccatgcc tgcctttact ctgccagagt tatattgctg gggttttgaa
     3061 gaagatccta ttaaataaaa gaataagcag tattattaag tagccctgca tttcaggttt
     3121 ccttgagtgg caggccaggc ctggccgtga acgttcactg aaatcatggc ctcttggcca
     3181 agattgatag cttgtgcctg tccctgagtc ccagtccatc acgagcagct ggtttctaag
     3241 atgctatttc ccgtataaag catgagaccg tgacttgcca gccccacaga gccccgccct
     3301 tgtccatcac tggcatctgg actccagcct gggttggggc aaagagggaa atgagatcat
     3361 gtcctaaccc tggaattgga tcctcttgtc ttacagatgg atctggagca acaaacttct
     3421 cactactcaa acaagcaggt gacgtggagg agaatcccgg ccccatgaaa tggaaagcac
     3481 tctttaccgc cgcaatcctt caagcacagt tgccaattac cgaggctgac attgtactga
     3541 cgcaaagtcc ccctagcttg gcgatgagtc tcgggaagcg agcgacgatt agttgccgag
     3601 cttctgaaag tgtcacaatc cttggctccc acctgatcca ttggtaccaa caaaaacctg
     3661 ggcagccccc gacgcttctc attcagttgg cgtctaacgt gcaaacagga gtaccggcca
     3721 gattttcagg ctcaggctct cgcaccgact ttactctgac catcgaccct gttgaggagg
     3781 atgatgtagc agtttactac tgtcttcaga gcagaaccat tcctcgcaca ttcggcggtg
     3841 gaacgaagtt ggaaatcaag ggctcaacaa gtgggagtgg gaagcccggc agcggggagg
     3901 gttctactaa aggccaaata cagttggttc aatccgggcc tgaactgaaa aagccgggag
     3961 agaccgtgaa aatttcttgc aaggctagcg ggtacacttt tacggattac tctattaact
     4021 gggttaagag ggcaccgggc aaagggctga aatggatggg ctggataaac accgagactc
     4081 gggagcctgc atatgcttat gatttcagag gtagatttgc gttctctttg gaaacctcag
     4141 cttcaacggc ctacttgcag attaataact tgaagtacga agacaccgcc acttacttct
     4201 gcgctctcga ttactcatac gctatggatt actggggcca gggcacgtcc gtgaccgtgt
     4261 ccagcgaaca gaaacttata tcagaggaag accttgcaat tgaagttatg tatcctcctc
     4321 cttacctaga caatgagaag agcaatggaa ccattatcca tgtgaaaggg aaacaccttt
     4381 gtccaagtcc cctatttccc ggaccttcta agcccttttg ggtgctggtg gtggttggtg
     4441 gagtcctggc ttgctatagc ttgctagtaa cagtggcctt tattattttc tgggtgagga
     4501 gtaagaggag caggctcctg cacagtgact acatgaacat gactccccgc cgccccgggc
     4561 ccacccgcaa gcattaccag ccctatgccc caccacgcga cttcgcagcc tatcgctcca
     4621 gagtgaagtt cagcaggagc gcagacgccc ccgcgtacca gcagggccag aaccagctct
     4681 ataacgagct caatctagga cgaagagagg agtacgatgt tttggacaag agacgtggcc
     4741 gggaccctga gatgggggga aagccgagaa ggaagaaccc tcaggaaggc ctgtacaatg
     4801 aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa ggcgagcgcc
     4861 ggaggggcaa ggggcacgat ggcctttacc agggtctcag tacagccacc aaggacacct
     4921 acgacgccct tcacatgcag gccctgcccc ctcgcggaag cggagctact aacttcagcc
     4981 tgctgaagca ggctggagac gtggaggaga accctggacc caatatccag aaccctgacc
     5041 ctgccgtgta ccagctgaga gactctaaat ccagtgacaa gtctgtctgc ctattcaccg
     5101 attttgattc tcaaacaaat gtgtcacaaa gtaaggattc tgatgtgtat atcacagaca
     5161 aaactgtgct agacatgagg tctatggact tcaagagcaa cagtgctgtg gcctggagca
     5221 acaaatctga ctttgcatgt gcaaacgcct tcaacaacag cattattcca gaagacacct
     5281 tcttccccag cccaggtaag ggcagctttg gtgccttcgc aggctgtttc cttgcttcag
     5341 gaatggccag gttctgccca gagctctggt caatgatgtc taaaactcct ctgattggtg
     5401 gtctcggcct tatccattgc caccaaaacc ctctttttac taagaaacag tgagccttgt
     5461 tctggcagtc cagagaatga cacgggaaaa aagcagatga agagaaggtg gcaggagagg
     5521 gcacgtggcc cagcctcagt ctctccaact gagttcctgc ctgcctgcct ttgctcagac
     5581 tgtttgcccc ttactgctct tctaggcctc attctaagcc ccttctccaa gttgacgcgt
     5641 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     5701 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag
     5761 cgcgcagctg cctgc
//
