LOCUS       Exported                4891 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  recombinant adeno-associated viral plasmid for delivery and 
            expression of ubiquitin variant that has reverted mutations L69P and
            V70L (i53-DM).
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-i53-DM
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4891
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   65..194
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        270..573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        574..777
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     728..748
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     774..798
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1378..1401
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1702..2178
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1901..1920)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2218..2358
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2218..2347
                     /label=AAV2 ITR
     rep_origin      2433..2888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2520..2539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2730..2751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2905..2924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3024..3046
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3084..3102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3170..3274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3275..4135
                     /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(3493..3512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4306..3
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4795..4814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
       61 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
      121 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      181 cactaggggt tcctgcggcc gcacgcgtct agttattaat agtaatcaat tacggggtca
      241 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      301 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      361 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      421 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      481 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      541 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      601 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      661 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      721 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      781 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      841 caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt gcattggaac
      901 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacaaa
      961 aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac tttccctaat
     1021 ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc attctaaaga
     1081 ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata aatatttctg
     1141 catataaatt gtaactgatg taagaggttt catattgcta atagcagcta caatccagct
     1201 accattctgc ttttatttta tggttgggat aaggctggat tattctgagt ccaagctagg
     1261 cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg ggcaacgtgc
     1321 tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat cgatatggac
     1381 tacaaagacg atgacgataa agccgccagt ttaaacggcg cgccattaat taaggatcca
     1441 atgttgattt tcgtgaaaac ccttaccggg aaaaccatca ccctcgaggt tgaaccctcg
     1501 gatacgatag aaaatgtaaa ggccaagatc caggataagg aaggaattcc tcctgatcag
     1561 cagagactgg cctttgctgg caaatcgctg gaagatggac gtactttgtc tgactacaat
     1621 attctaaagg actctaaact tcatctagtg ttgagacttc gttgaaagct tgcctcgagc
     1681 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     1741 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     1801 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     1861 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     1921 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     1981 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     2041 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     2101 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     2161 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     2221 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2281 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2341 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2401 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2461 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2521 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2581 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2641 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2701 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2761 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     2821 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2881 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2941 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3001 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3061 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3121 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4141 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4201 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4441 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4861 gtgatgctcg tcaggggggc ggagcctatg g
//
