LOCUS       sequence-275596-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA1 E638K 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 E638K
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        51..299
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        319..394
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        408..656
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        677..752
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(855..1443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(935..954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1614..2405,2406..2474))
                     /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     2237..2256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2475..2579)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaaggaacc aattcagtcg actggatccg gtaccaaggt cgggcaggaa gagggcctat
       61 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
      121 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
      181 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
      241 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
      301 agacgattaa tgcgtctcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      361 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt taagcttgag ggcctatttc
      421 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
      481 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
      541 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
      601 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcaa
      661 ttcagtacaa ttaggtgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      721 atcaacttga aaaagtggca ccgagtcggt gctttttttc tagagaattc ctcgaggtac
      781 ctctctacat atgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      841 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      901 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      961 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1021 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1081 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1141 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1201 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1261 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1321 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1381 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1441 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1501 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1561 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1621 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1681 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1741 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1801 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1861 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1921 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1981 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2041 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2101 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2161 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2221 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2281 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2341 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2401 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2461 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2521 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2581 acatttcccc gaaaagtgcc acctgacgtc gctagctgta caaaaaagca ggctt
//
