LOCUS       sequence-275581-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA2 V572I using base 
            editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2635)
  AUTHORS   Billon P, Nambiar TS, Hayward SB, Zafra MP, Schatoff EM, Oshima K, 
            Dunbar A, Breinig M, Park YC, Ryu HS, Tschaharganeh DF, Levine RL, 
            Baer R, Ferrando A, Dow LE, Ciccia A
  TITLE     Detection of Marker-Free Precision Genome Editing and Genetic 
            Variation through the Capture of Genomic Signatures.
  JOURNAL   Cell Rep. 2020 Mar 10;30(10):3280-3295.e6. doi: 
            10.1016/j.celrep.2020.02.068.
  PUBMED    32160537
REFERENCE   2  (bases 1 to 2635)
  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: 
            "10.1016/j.celrep.2020.02"; journalName: "Cell Rep"; date: 
            "2020-03-10- 10"; volume: "30"; issue: "10"; pages: "3280-3295.e6"
COMMENT     Sequence Label: sgRNA BRCA2 V572I
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..298
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        318..393
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        407..655
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        676..751
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(854..1442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(934..953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1613..2404,2405..2473))
                     /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     2236..2255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2474..2578)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaggaacca attcagtcga ctggatccgg taccaaggtc gggcaggaag agggcctatt
       61 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat
      121 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      181 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      241 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccga
      301 gacgattaat gcgtctcgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      361 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt aagcttgagg gcctatttcc
      421 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa
      481 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
      541 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
      601 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggcta
      661 cagaattctg tgtgggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      721 tcaacttgaa aaagtggcac cgagtcggtg ctttttttct agagaattcc tcgaggtacc
      781 tctctacata tgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      841 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      901 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      961 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1021 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1081 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1141 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1201 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1261 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1321 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1381 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1441 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1501 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1561 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1621 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1681 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1741 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1801 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1861 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1921 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1981 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2041 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2101 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2161 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2221 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2281 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2341 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2401 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2461 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2581 catttccccg aaaagtgcca cctgacgtcg ctagctgtac aaaaaagcag gcttt
//
