LOCUS       sequence-275583-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA2 V2102I 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 V2102I
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        330..405
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        419..667
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        688..763
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(866..1454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(946..965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1625..2416,2417..2485))
                     /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     2248..2267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2486..2590)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagcaggct ttaaaggaac caattcagtc gactggatcc ggtaccaagg tcgggcagga
       61 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      121 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      301 acgaaacacc gagacgatta atgcgtctcg ttttagagct agaaatagca agttaaaata
      361 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttaagcttga
      421 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
      481 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
      541 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
      601 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
      661 aaacaccgat acattttgtc tagacgtgtt ttagagctag aaatagcaag ttaaaataag
      721 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctagagaatt
      781 cctcgaggta cctctctaca tatgacatgt gagcaaaagg ccagcaaaag gccaggaacc
      841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1381 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1441 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1501 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1561 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1621 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1681 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1741 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1801 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1861 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1921 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1981 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2041 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2101 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2161 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2221 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2281 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2341 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2401 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2461 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2521 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2581 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgctagctgt acaaa
//
