LOCUS       sequence-199153-        2962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning of guide RNAs compatible with RanCas13b. Contains a 3' 
            direct repeat. Clone using BbsI (BpiI). F overhang cacc. R overhang 
            caac..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2962)
  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 2962)
  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: pC0041-RanCas13b crRNA backbone
FEATURES             Location/Qualifiers
     source          1..2962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(9..14,15..32,33..39)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    47..63
                     /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     52..74
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     71..87
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     71..87
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        101..349
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     101..121
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     272..291
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(433..450)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(433..449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(442..464)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(658..677)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     777..799
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(837..855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        923..1027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1028..1096,1097..1888)
                     /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(1246..1265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2059..2647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2548..2567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2801..2818
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2935..2956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
       61 caatttcaca caggaaacag ctatgaccat gattacgcca gagggcctat ttcccatgat
      121 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
      181 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
      241 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
      301 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacacca tgtcttcctg
      361 ggacgaagac aagttgggac tgctctcact ttgaagggta ttcacaactt ttttggtacc
      421 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      481 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      541 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      601 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      661 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      721 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      781 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      841 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      901 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      961 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1021 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1081 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1141 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1201 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1261 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1321 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1381 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1441 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1501 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1561 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1621 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1681 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1741 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1801 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1861 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1921 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2161 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2281 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2341 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2401 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2461 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2521 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2581 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2641 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2701 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2761 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2821 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2881 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2941 gagttagctc actcattagg ca
//
