LOCUS       sequence-199148-        2990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Guide RNA expression plasmid for dPspCas13b that can be used to 
            direct ADAR deaminase domain activity to the W85X mutation in the 
            luciferase reporter plasmid pC0038.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2990)
  AUTHORS   Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung 
            J, Zhang F
  TITLE     RNA editing with CRISPR-Cas13.
  JOURNAL   Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
  PUBMED    29070703
REFERENCE   2  (bases 1 to 2990)
  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: "Science. 
            2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180."
COMMENT     Sequence Label: pC0051-W85X REPAIR targeting guide cloned into...
FEATURES             Location/Qualifiers
     source          1..2990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      479..1067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     968..987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1221..1238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1355..1376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1391..1396,1397..1414,1415..1421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1429..1445
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1453..1469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1453..1469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1483..1731
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1483..1503
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1654..1673
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1843..1860)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1843..1859)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1852..1874)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2068..2087)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2187..2209
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2247..2265)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2333..2437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2438..2506,2507..2990,1..308)
                     /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     complement(2656..2675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
       61 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      121 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      181 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      241 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      301 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      361 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1141 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1201 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     1261 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1321 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     1381 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     1441 aacaatttca cgcaggaaac agctatgacc atgattacgc cagagggcct atttcccatg
     1501 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
     1561 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     1621 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     1681 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cgatagaatg
     1741 ttctaaacca tcctgcggcc tctactctgc attcaattac atgttgtgga aggtccagtt
     1801 ttgaggggct attacaactt ttttggtacc gagctcgaat tcactggccg tcgttttaca
     1861 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     1921 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1981 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2041 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2101 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2161 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2221 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     2281 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
//
