LOCUS       sequence-275604-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA1 E1754K 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 E1754K
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        43..291
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        311..386
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        400..648
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        669..744
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(847..1435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(927..946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1606..2397,2398..2466))
                     /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     2229..2248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2467..2571)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaattcagt cgactggatc cggtaccaag gtcgggcagg aagagggcct atttcccatg
       61 attccttcat atttgcatat acgatacaag gctgttagag agataattag aattaatttg
      121 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
      181 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
      241 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cgagacgatt
      301 aatgcgtctc gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      361 ttgaaaaagt ggcaccgagt cggtgctttt tttaagcttg agggcctatt tcccatgatt
      421 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
      481 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      541 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
      601 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg ggattctctt
      661 gctcgcttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      721 gaaaaagtgg caccgagtcg gtgctttttt tctagagaat tcctcgaggt acctctctac
      781 atatgacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2581 ccgaaaagtg ccacctgacg tcgctagctg tacaaaaaag caggctttaa aggaa
//
