LOCUS       Exported                2635 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Plasmid expressing a sgRNA to introduce BRCA2 E2772K using base 
            editing.
ACCESSION   .
VERSION     .
KEYWORDS    sgRNA BRCA2 E2772K
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2635
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        57..297
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        325..400
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        414..654
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        683..758
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(861..1449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(941..960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1620..2480)
                     /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     2243..2262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2481..2585)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aggctttaaa ggaaccaatt cagtcgactg gatccggtac caaggtcggg caggaagagg
       61 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
      121 ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
      181 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
      241 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
      301 acaccgagac gattaatgcg tctcgtttta gagctagaaa tagcaagtta aaataaggct
      361 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttaag cttgagggcc
      421 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg
      481 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
      541 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
      601 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
      661 ccggagattc tggggcttca aggttttaga gctagaaata gcaagttaaa ataaggctag
      721 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctaga gaattcctcg
      781 aggtacctct ctacatatga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      841 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      901 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      961 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1021 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1081 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1141 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1201 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1261 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1321 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1381 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1441 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1501 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1561 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1621 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1681 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1741 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1801 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1861 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1921 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1981 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2041 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2101 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2161 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2221 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2281 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2341 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2401 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2461 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2521 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2581 ccgcgcacat ttccccgaaa agtgccacct gacgtcgcta gctgtacaaa aaagc
//
