LOCUS       sequence-182617-        4891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recombinant adeno-associated viral plasmid for delivery and 
            expression of ubiquitin variant (i53) that is a selective inhibitor 
            of 53BP1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4891)
  AUTHORS   Canny MD, Moatti N, Wan LCK, Fradet-Turcotte A, Krasner D, 
            Mateos-Gomez PA, Zimmermann M, Orthwein A, Juang YC, Zhang W, 
            Noordermeer SM, Seclen E, Wilson MD, Vorobyov A, Munro M, Ernst A, 
            Ng TF, Cho T, Cannon PM, Sidhu SS, Sicheri F, Durocher D
  TITLE     Inhibition of 53BP1 favors homology-dependent DNA repair and 
            increases CRISPR-Cas9 genome-editing efficiency.
  JOURNAL   Nat Biotechnol. 2018 Jan;36(1):95-102. doi: 10.1038/nbt.4021. Epub 
            2017 Nov 27.
  PUBMED    29176614
REFERENCE   2  (bases 1 to 4891)
  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"; doi: "10.1038/nbt.4021"; 
            journalName: "Nat Biotechnol"; date: "2018-01"; volume: "36"; issue:
            "1"; pages: "95-102"
COMMENT     Sequence Label: pAAV-i53
FEATURES             Location/Qualifiers
     source          1..4891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   58..187
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        263..566
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        567..770
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     721..741
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     767..791
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          919..1297
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             1371..1394
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1695..2171
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1894..1913)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2211..2351
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2211..2340
                     /label=AAV2 ITR
     rep_origin      2426..2881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2513..2532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2723..2744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2898..2917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3017..3039
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3077..3095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3163..3267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3268..3336,3337..4128)
                     /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     complement(3486..3505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4299..4887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4788..4807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
       61 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
      121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      181 ggttcctgcg gccgcacgcg tctagttatt aatagtaatc aattacgggg tcattagttc
      241 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      301 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      361 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      421 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      481 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      541 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      601 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      661 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      721 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      781 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
      841 accgatccag cctccgcgga ttcgaatccc ggccgggaac ggtgcattgg aacgcggatt
      901 ccccgtgcca agagtgacgt aagtaccgcc tatagagtct ataggcccac aaaaaatgct
      961 ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct aatctctttc
     1021 tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa agaataacag
     1081 tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt ctgcatataa
     1141 attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc
     1201 tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct aggccctttt
     1261 gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg tgctggtctg
     1321 tgtgctggcc catcactttg gcaaagaatt gggattcgaa catcgatatg gactacaaag
     1381 acgatgacga taaagccgcc agtttaaacg gcgcgccatt aattaaggat ccaatgttga
     1441 ttttcgtgaa aacccttacc gggaaaacca tcaccctcga ggttgaaccc tcggatacga
     1501 tagaaaatgt aaaggccaag atccaggata aggaaggaat tcctcctgat cagcagagac
     1561 tggcctttgc tggcaaatcg ctggaagatg gacgtacttt gtctgactac aatattctaa
     1621 aggactctaa acttcatcct ctgttgagac ttcgttgaaa gcttgcctcg agcagcgctg
     1681 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     1741 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     1801 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     1861 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     1921 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     1981 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     2041 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     2101 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     2161 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     2221 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2281 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2341 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2401 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2461 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     2521 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2581 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2641 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2701 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     2761 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     2821 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     2881 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2941 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3001 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3061 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3121 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3241 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3301 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3361 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3421 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3481 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3541 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3601 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3661 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3721 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3781 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3841 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3901 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3961 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4021 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4081 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4141 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4201 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4261 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4381 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4441 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4501 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4561 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4621 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4681 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4741 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4801 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4861 tcgtcagggg ggcggagcct atggaaaaac g
//
