LOCUS       sequence-190286-        2634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  B52 plasmid expressing ATP1A1 sgSTOP (cloned in BsmBI site) and 
            containing an empty sgRNA-expression cassette (use BbsI for 
            cloning).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2634)
  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 2634)
  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 (plasmid expressing ATP1A1 sgSTOP and
            con...
FEATURES             Location/Qualifiers
     source          1..2634
                     /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        707..782
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(885..1473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(965..984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1644..2435,2436..2504))
                     /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     2267..2286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2505..2609)
                     /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 gaaacaccgg gtcttcgaga agacctgttt tagagctaga
      721 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      781 gctttttttc tagagaattc ctcgaggtac ctctctacat atgacatgtg agcaaaaggc
      841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct
//
