LOCUS       sequence-199182-        2970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-targeting guide RNA for use with PspCas13b. Useful as a control 
            for RNA editing and RNA knockdown experiments..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2970)
  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 2970)
  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: pC0052-REPAIR non-targeting guide clone into p...
FEATURES             Location/Qualifiers
     source          1..2970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..129
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    246..267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(282..287,288..305,306..312)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    320..336
                     /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     344..360
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     344..360
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        374..622
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     374..394
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     545..564
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(714..731)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(714..730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(723..745)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(939..958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1058..1080
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1118..1136)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1204..1308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1309..1377,1378..2169)
                     /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(1527..1546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2340..2928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2829..2848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      121 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acgcaggaaa cagctatgac
      361 catgattacg ccagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
      421 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
      481 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      541 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      601 cttgtggaaa ggacgaaaca ccggtaatgc ctggcttgtc gacgcatagt ctggttgtgg
      661 aaggtccagt tttgaggggc tattacaact tttttggtac cgagctcgaa ttcactggcc
      721 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
      781 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
      841 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
      901 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
      961 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     1021 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     1081 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     1141 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     1201 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1261 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1321 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1381 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1441 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1501 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1561 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1621 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1681 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1741 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1801 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1861 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1921 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1981 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2041 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2101 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2161 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2461 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2941 gcggcctttt tacggttcct ggccttttgc
//
