LOCUS       sequence-443109-        2244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hU6-driven expression of CasRx guide RNA targeting EZH2. 5' 
            processed DR followed by EZH2 guide..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2244)
  AUTHORS   Yan H, Tang W
  TITLE     Programmed RNA editing with an evolved bacterial adenosine 
            deaminase.
  JOURNAL   Nat Chem Biol. 2024 Jul 5. doi: 10.1038/s41589-024-01661-x.
  PUBMED    38969862
REFERENCE   2  (bases 1 to 2244)
  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 Chem 
            Biol. 2024 Jul 5. doi: 10.1038/s41589-024-01661-x."
COMMENT     Sequence Label: CasRx gRNA targeting EZH2
FEATURES             Location/Qualifiers
     source          1..2244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     371..390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(609..713)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        820..1068
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     820..840
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     991..1010
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1070..1099
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     rep_origin      complement(1233..1821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1313..1332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1992..2244,1..539,540..608))
                     /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"
ORIGIN
        1 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
       61 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
      241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
      301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
      361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
      421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      721 ccccgaaaag tgccacctga cgtcgctagc tgtacaaaaa agcaggcttt aaaggaacca
      781 attcagtcga ctggatccgg taccaaggtc gggcaggaag agggcctatt tcccatgatt
      841 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
      901 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      961 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     1021 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccga acccctacca
     1081 actggtcggg gtttgaaacg agagcagcag caaactcctt tgctccctct ttttttaagc
     1141 ttgggccgct cgaggtacct ctctacatat gacatgtgag caaaaggcca gcaaaaggcc
     1201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2221 gcctccatcc agtctattaa ttgt
//
