LOCUS       sequence-190284-        2637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  B270 plasmid expressing SMARCAL1 sgSTOP (cloned in BbsI site) in 
            combination with ATP1A1 sgSTOP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2637)
  AUTHORS   Billon P, Bryant EE, Joseph SA, Nambiar TS, Hayward SB, Rothstein R,
            Ciccia A
  TITLE     CRISPR-Mediated Base Editing Enables Efficient Disruption of 
            Eukaryotic Genes through Induction of STOP Codons.
  JOURNAL   Mol Cell. 2017 Sep 4. pii: S1097-2765(17)30605-6. doi: 
            10.1016/j.molcel.2017.08.008.
  PUBMED    28890334
REFERENCE   2  (bases 1 to 2637)
  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"; journalName: "Mol Cell. 
            2017 Sep 4. pii: S1097-2765(17)30605-6. doi: 
            10.1016/j.molcel.2017.08.008."
COMMENT     Sequence Label: B270 + SMARCAL1 sgSTOP
FEATURES             Location/Qualifiers
     source          1..2637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        351..426
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        440..688
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        710..785
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(888..1476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(968..987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1647..2438,2439..2507))
                     /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     2270..2289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2508..2612)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gatccaagct gctacagaag gttttagagc
      361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      421 cggtgctttt tttaagcttg agggcctatt tcccatgatt ccttcatatt tgcatatacg
      481 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
      541 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      601 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      661 atatatcttg tggaaaggac gaaacaccgc agcatcagag gactagctcg ttttagagct
      721 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      781 ggtgcttttt ttctagagaa ttcctcgagg tacctctcta catatgacat gtgagcaaaa
      841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1321 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacct
//
