LOCUS       sequence-275608-        2636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a sgRNA to introduce FANCD2 Q223Stop using base 
            editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2636)
  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 2636)
  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 FANCD2 Q223Stop
FEATURES             Location/Qualifiers
     source          1..2636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..300
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        320..395
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        409..657
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        679..754
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(857..1445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(937..956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1616..2407,2408..2476))
                     /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     2239..2258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2477..2581)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaaaggaac caattcagtc gactggatcc ggtaccaagg tcgggcagga agagggccta
       61 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga
      121 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
      181 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
      241 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
      301 gagacgatta atgcgtctcg ttttagagct agaaatagca agttaaaata aggctagtcc
      361 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttaagcttga gggcctattt
      421 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
      481 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
      541 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
      601 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgat
      661 tcccagcacg ctgatgtggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      721 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tctagagaat tcctcgaggt
      781 acctctctac atatgacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1261 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2581 gcacatttcc ccgaaaagtg ccacctgacg tcgctagctg tacaaaaaag caggct
//
