LOCUS       sequence-354677-        1882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUC19 derivative plasmid used as a substrate for SpRYgest, 
            restriction enzyme, and TtAgo digests. Encodes an SpCas9 
            EMX1-site1-NGG-PAM target site between NheI and HindIII restriction 
            sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1882)
  AUTHORS   Christie KA, Guo JA, Silverstein RA, Doll RM, Mabuchi M, Stutzman 
            HE, Lin J, Ma L, Walton RT, Pinello L, Robb GB, Kleinstiver BP
  TITLE     Precise DNA cleavage using CRISPR-SpRYgests.
  JOURNAL   Nat Biotechnol. 2022 Oct 6. pii: 10.1038/s41587-022-01492-y. doi: 
            10.1038/s41587-022-01492-y.
  PUBMED    36203014
REFERENCE   2  (bases 1 to 1882)
  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: "Nat 
            Biotechnol. 2022 Oct 6. pii: 10.1038/s41587-022-01492-y. doi: 
            10.1038/s41587-022-01492-y."
COMMENT     Sequence Label: pUC19-SpRYgest_targetplasmid (KAC833)
FEATURES             Location/Qualifiers
     source          1..1882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(140..728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(220..239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(899..1690,1691..1759))
                     /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     1522..1541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1760..1864)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctgac gtcgctagcg agtccgagca gaagaagaag ggcaagcttg ggccgctcga
       61 ggtacctctc tacatatgac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
      961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1861 cgcgcacatt tccccgaaaa gt
//
