LOCUS       sequence-189661-        7677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CgCas13a for bacterial expression and contains backbone 
            for spacer cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7677)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, 
            Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, 
            Voytas DF, Ting AY, Zhang F
  TITLE     RNA targeting with CRISPR-Cas13.
  JOURNAL   Nature. 2017 Oct 4. doi: 10.1038/nature24049.
  PUBMED    28976959
REFERENCE   2  (bases 1 to 7677)
  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: "Nature. 
            2017 Oct 4. doi: 10.1038/nature24049."
COMMENT     Sequence Label: pC024 - CgCas13a from Carnobacterium gallinaru...
FEATURES             Location/Qualifiers
     source          1..7677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(167..201)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        complement(join(3570..3575,3576..3593,3594..3599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      3637..4182
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4083..4102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4299..4316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4708..4810
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4811..5470
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4877..4896)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(join(6504..7677,1..17))
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     7323..7342
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
       61 ccgcattaaa gcttgttata gtcctcttac atttagaggt agtctttaat cgagacgaat
      121 ctccgtctcc gttatagtcc tcttacattt agaggtagtc tttaatgcta gcattatacc
      181 taggactgag ctagctgtca atcagccgtg cttgtccagg atcttgatca tagacttggt
      241 cacggcgttc ttcagctttc tatcgtaggc catcaggtcc cgcagattgt tcatgctctc
      301 gatcatagac agctccagct gcagcacgga caggtgggct gtctgatttc tgatcttgcc
      361 gtgcacggcc ttgatgccga tcacgttctc cagataggcg tgcagccgat ccaggaaggg
      421 cttcacctct ggctcattgc ccttgtaagg attctcctcg ttgtcgatca ctttgatgtt
      481 cttgatcttg atgggcttca cctcggcatt cagccgcagc acgccggaca ggttcttata
      541 atcgctgatg tggtatgtct cgccgcccag ctcagaaaag gagcggctga agtacaccag
      601 atccctctca aacagggcgc tgaagcccac cattctgccc agcaggtctg tgatcagata
      661 gtgcagctga gacaggtatg tcagggtcac cttattggcc ttccagttga accgatcgat
      721 cttcttacaa tagtctctgt actcctcccg cttctctgct cccagtgtct gctgtccctc
      781 tgcccactcg gtgtgcagat tcttgcgctt ctccaccagc tcgccgatct cctccctctt
      841 gtgcatccac tcctggatat cggtggcggc cacgcgaaac tgtgggttgt ccttcagcag
      901 ggcctcgatc aggctctttg tgtggtactt ctccagcttc aggattggcc tgaaagacac
      961 gcgtgtccta tcgtcggtct gcacataagg ggctgtctcg tacaggctct tatcctcaaa
     1021 ataggcgctc acgtccacat tctgggagtc gttcttctca tcgtagtcgt tagacaggat
     1081 ggtcagctcc acgataggca gcagattctg gatcagctcg gcgtgctgtg tgtggttgaa
     1141 cttgatccgg ctctgcttga acttgatcat ctcatttctc agctcgttca ggtgccggga
     1201 gtccagcagc ttgcagaaca gccagtagct gatggcagaa ttggtggcca ccagctggtg
     1261 ctccacctgg atgctggcct tcagggtttt ctcccgctcg ttgatcttct tagacaggga
     1321 gatctctgtc tccttgatca ctttctcctc ggagctcagc agcagctcct tgccggcaaa
     1381 tgtggtcagg aacacgtcga agcccttcac gaagatctgc atcagcagct tctcaaaatt
     1441 catggtgatg ttcttctcga agcccttatc gttcttctcg gccttgcgca ccttctcctc
     1501 cctcatctcg gactgcagat agctcatgta ctggtcgggt gtctccttga actccatctc
     1561 gcgcacctgc tcaaaggcgt acttcctcag gaacttctta ttcttcttca tcttctttct
     1621 ggcctcgtcc ttgttgatcc gcaggatggc cttcactgtc tcgcggaagg ctggagaatt
     1681 gccctgggaa aagttaggca ggaacacgta ctggtagatc agcttcagca gatagtactg
     1741 ggcctgaaaa gactcctcct tcagctcata cctattatag taggacaggc ccaggtagtt
     1801 ggcggtgccc tctgtggcgg tctgatagga gtggcccttc ttcaccagct tcttgtagct
     1861 aggggcgaag gccaccacct gggagcccag gctaaacttc atctccttct tggcgaacag
     1921 cttctccagg gacttcaggg gatagtactc cagggcgcca gaggacttga tgcgctcgat
     1981 aaagaactgc tccagctggt tcttctcggc ctcaaacagg aacttataat ctgtctcgat
     2041 gtagctctga gacttgccgg tgctgttctc ggaggcatca aactcgaagt tgctcttgtc
     2101 cagctggggc agattgaact tctggttctt gtcaaagctg tggaagatct cattgcggat
     2161 ctgctgcaca gagcccctca gggcccagat gttctgcatc tgcttggcct cggtctgctc
     2221 gccaaatggg atcagctcgg actccctatc cttcatgccc ttcagctcaa aagggaaata
     2281 gggaaagaag tggtggtaat tcacgaagct agatgtggcg atggcctcct taaactctct
     2341 cttgcccagg atgtcgccgc tccgcatagg ctcgatgatg ttcttcaggg agcttgtggc
     2401 gaacacgcag gcatccataa acttggtctg gaagccctca tagatgccga tctcctgcag
     2461 cttcttggag tccagcacct gattctccag gccaaactgc ttatacttgc cgatctgcat
     2521 caggtaggag atcagggcat tcttcagctg gtagttcact gtggtggcga ttgtctcgct
     2581 gttcagatag tagggctcat cctcggtgca tctctccttc ttgattggaa aatagtgctc
     2641 gaagtacttg ccgatctcga tattgaactg gttcagctcc tcattctcct tgatcttctc
     2701 gatcaggccg atgtgtgtct tatatgggta gtccttccag atggtcttga tcttaaagtt
     2761 cttctgcttc ttataggcct ccttctcctt ctcaggattc ttgcccttct tcatgctctt
     2821 ctccagatcg ttcagctcgt cctgcactct cttgcacatc ttcttgaagt tgtagatctc
     2881 ggccagctta gacagcttct catccaggat ggaaaagtcg cccctctcct gcacctcgat
     2941 tgcccactgg tgcagcactc tcttccgctt gctgaatgtc tcgggattga tagagaagcc
     3001 cttattctgc tcgatggagt tggccagcag gatgatcttt ctctcgatga acttgttctt
     3061 atactcccgg tagggcatca tctcctggca aaaatcggcc caagactcgt cggtgaagtg
     3121 ctcgcctggc accttctgca gggcattata aaacagggag atcagatcca gattcttgat
     3181 ctccacgata tcgtcgttct taaacttgcg cttgatgaac agggccaggt ccttcttagg
     3241 cagatccttc aggtctgtca gggacatctt gggcttcctg atattcttgt tggtatccac
     3301 gcccaccagc ttctcgaaga tctggctgat gtagtcgccg ttcttcttga ttgtggcatt
     3361 cttgttctcc ttctgggaat tgatggtctt gctgtgaaaa gacttgttga agctggcctt
     3421 cttcaggcgc aggatctcgg cggtggactt cctgctctcc tcattcagct tcagtgtgcc
     3481 atacttctcc tccttctgca tggtcacctg gtacagcttg ttgtccagct tgatcttcac
     3541 ctttgtgatc cgcatggata agacctccta acatacgagc cggaagcata aagtgtaaat
     3601 ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt
     3661 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
     3721 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
     3781 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
     3841 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
     3901 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
     3961 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
     4021 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
     4081 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
     4141 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
     4201 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
     4261 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
     4321 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
     4381 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
     4441 cgctagcgct gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc agtagctgaa
     4501 caggagggac agctgataga aacagaagcc actggagcac ctcaaaaaca ccatcataca
     4561 ctaaatcagt aagttggcag catcacccga cgcactttgc gccgaataaa tacctgtgac
     4621 ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg
     4681 ggccaacttt tggcgaaaat gacacgttga tcggcacgta agaggttcca actttcacca
     4741 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     4801 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     4861 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     4921 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     4981 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
     5041 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     5101 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     5161 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     5221 gtttattgag aatatgtttt tcgtgtcagc caatccctgg gtgagtttca ccagttttga
     5281 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     5341 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     5401 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     5461 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     5521 aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc
     5581 ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg
     5641 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     5701 ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca gagcctgata
     5761 aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     5821 ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg ggtatggtgg
     5881 caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg agttgcatga
     5941 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     6001 taccggcagc ggtgcggact gttgtaactc agaataagaa atgaggccgc tcatggcgtt
     6061 gactctcagt catagtatcg tggtatcacc ggttggttcc actctctgtt gcgggcaact
     6121 tcagcagcac gtaggggact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     6181 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     6241 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     6301 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
     6361 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
     6421 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     6481 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     6541 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     6601 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     6661 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     6721 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     6781 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     6841 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     6901 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     6961 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     7021 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     7081 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta
     7141 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     7201 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     7261 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     7321 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     7381 acgatgcgtc cggcgtagag gatccacagg acgggtgtgg tcgccatgat cgcgtagtcg
     7441 atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt
     7501 gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca
     7561 tagtgactgg cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc
     7621 ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagc
//
