LOCUS       sequence-275599-        2635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA1 T922I 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 T922I
FEATURES             Location/Qualifiers
     source          1..2635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..309
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        329..404
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        418..666
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        687..762
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(865..1453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(945..964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1624..2415,2416..2484))
                     /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     2247..2266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2485..2589)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagcaggctt taaaggaacc aattcagtcg actggatccg gtaccaaggt cgggcaggaa
       61 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
      121 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      181 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      241 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      301 cgaaacaccg agacgattaa tgcgtctcgt tttagagcta gaaatagcaa gttaaaataa
      361 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt taagcttgag
      421 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
      481 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
      541 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
      601 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
      661 aacaccgaga cagttaatat cactgcgttt tagagctaga aatagcaagt taaaataagg
      721 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc tagagaattc
      781 ctcgaggtac ctctctacat atgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1921 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2581 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gctagctgta caaaa
//
