LOCUS       sequence-275595-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA1 C64Y 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 C64Y
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..297
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        317..392
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        406..654
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        675..750
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(853..1441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(933..952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1612..2403,2404..2472))
                     /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     2235..2254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2473..2577)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaggaaccaa ttcagtcgac tggatccggt accaaggtcg ggcaggaaga gggcctattt
       61 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt
      121 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
      181 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
      241 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgag
      301 acgattaatg cgtctcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      361 atcaacttga aaaagtggca ccgagtcggt gcttttttta agcttgaggg cctatttccc
      421 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
      481 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
      541 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
      601 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgtacat
      661 aaaggacact gtgagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
      721 caacttgaaa aagtggcacc gagtcggtgc tttttttcta gagaattcct cgaggtacct
      781 ctctacatat gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1021 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1261 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2521 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2581 atttccccga aaagtgccac ctgacgtcgc tagctgtaca aaaaagcagg cttta
//
