LOCUS       sequence-275607-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA1 Q1779Stop 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 BRCA1 Q1779Stop
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..296
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        316..391
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        405..653
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        674..749
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(852..1440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(932..951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1611..2402,2403..2471))
                     /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     2234..2253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2472..2576)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aggaaccaat tcagtcgact ggatccggta ccaaggtcgg gcaggaagag ggcctatttc
       61 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attagaatta
      121 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
      181 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
      241 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgaga
      301 cgattaatgc gtctcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      361 tcaacttgaa aaagtggcac cgagtcggtg ctttttttaa gcttgagggc ctatttccca
      421 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
      481 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
      541 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
      601 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgttagat
      661 caactggaat ggagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      721 aacttgaaaa agtggcaccg agtcggtgct ttttttctag agaattcctc gaggtacctc
      781 tctacatatg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1261 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2581 tttccccgaa aagtgccacc tgacgtcgct agctgtacaa aaaagcaggc tttaa
//
