LOCUS       sequence-181267-        8479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CaCas13a 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 8479)
  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 8479)
  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: pC023 - CaCas13a from [Clostridium] aminophilu...
FEATURES             Location/Qualifiers
     source          1..8479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        228..262
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             410..1600
                     /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     complement(762..781)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(2634..3293)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3208..3227
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3294..3396)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3788..3805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3922..4467)
                     /direction=LEFT
                     /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     complement(4002..4021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(4505..4510,4511..4528,4529..4534)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 atctgaacac cgtggcccgc ctgctgtact atccacacga ggccaagaag agcatcgtgg
       61 acacacggga gaagaagaac aacaagaaaa ccaatagagg cgatggcaca ttcaacaagc
      121 agaagggcac cgccaggaag gagaaggaca atggcccccg cgagtttaac gataccggct
      181 tctccaatac accatttgcc ggcttcgatc cctttagaaa ctcttgattg acagctagct
      241 cagtcctagg tataatgcta gcgtctattg ccctctatat cgggctgttc tccaaacgga
      301 gacggagatt cgtctcggtc tattgccctc tatatcgggc tgttctccaa acaagcttta
      361 atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa
      421 caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt
      481 tatgccggta ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca
      541 ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg
      601 agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac
      661 tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atcctctacg ccggacgcat
      721 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
      781 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
      841 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
      901 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
      961 gcataaggga gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     1021 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     1081 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     1141 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     1201 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     1261 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     1321 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     1381 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     1441 cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     1501 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     1561 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     1621 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     1681 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     1741 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     1801 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     1861 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     1921 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcccc tacgtgctgc
     1981 tgaagttgcc cgcaacagag agtggaacca accggtgata ccacgatact atgactgaga
     2041 gtcaacgcca tgagcggcct catttcttat tctgagttac aacagtccgc accgctgccg
     2101 gtagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2161 ttatcatgca actcgtagga caggtgccgg cagcgcccaa cagtcccccg gccacggggc
     2221 ctgccaccat acccacgccg aaacaagcgc cctgcaccat tatgttccgg atctgcatcg
     2281 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctaaccgt
     2341 ttttatcagg ctctgggagg cagaataaat gatcatatcg tcaattatta cctccacggg
     2401 gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     2461 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     2521 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt
     2581 attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc
     2641 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     2701 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     2761 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     2821 aaatcaaaac tggtgaaact cacccaggga ttggctgaca cgaaaaacat attctcaata
     2881 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     2941 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     3001 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     3061 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     3121 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     3181 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     3241 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     3301 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     3361 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt gtcattttcg ccaaaagttg
     3421 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     3481 tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt
     3541 tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc
     3601 ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta
     3661 gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat
     3721 gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt
     3781 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     3841 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     3901 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     3961 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4021 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     4081 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     4141 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     4201 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     4261 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     4321 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     4381 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     4441 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     4501 tagatttaca ctttatgctt ccggctcgta tgttaggagg tcttatccat gaagttctct
     4561 aaggtggacc acacccggag cgccgtgggc atccagaagg ccaccgactc tgtgcacggc
     4621 atgctgtata ccgaccccaa gaagcaggaa gtgaatgatc tggacaagag gtttgatcag
     4681 ctgaacgtga aggccaagcg cctgtataac gtgttcaatc agtccaaggc cgaggaggac
     4741 gatgacgaga agcggtttgg caaggtggtg aagaagctga acagagagct gaaggacctg
     4801 ctgttccacc gggaggtgag cagatacaac tccatcggca atgccaagta taactactat
     4861 ggcatcaagt ccaaccctga ggagatcgtg tctaatctgg gcatggtgga gagcctgaag
     4921 ggagagcggg acccccagaa agtgatcagc aagctgctgc tgtactatct gagaaagggc
     4981 ctgaagcctg gcaccgacgg cctgaggatg atcctggagg cctcctgcgg cctgcgcaag
     5041 ctgtctggcg atgagaagga gctgaaggtg ttcctgcaga ccctggatga ggactttgag
     5101 aagaaaacct tcaagaagaa cctgatccgg tctatcgaga accagaatat ggccgtgcag
     5161 ccaagcaacg agggcgaccc catcatcggc atcacccagg gccggtttaa tagccagaag
     5221 aacgaggaga agtccgccat cgagagaatg atgtctatgt acgccgatct gaacgaggac
     5281 cacagggagg atgtgctgcg caagctgcgg agactgaatg tgctgtattt caacgtggac
     5341 accgagaaaa ccgaggagcc caccctgcct ggagaggtgg atacaaatcc agtgtttgaa
     5401 gtgtggcacg accacgagaa gggcaaggag aacgatagac agtttgccac cttcgccaag
     5461 atcctgaccg aggacaggga gacacgcaag aaggagaagc tggccgtgaa ggaggccctg
     5521 aatgacctga agagcgccat cagggatcac aacatcatgg cctaccgctg ttccatcaag
     5581 gtgaccgagc aggataagga cggcctgttc tttgaggacc agcggatcaa tagattctgg
     5641 attcaccaca tcgagtctgc cgtggagaga atcctggcca gcatcaaccc cgagaagctg
     5701 tataagctgc gcatcggcta tctgggcgag aaagtgtgga aggatctgct gaattacctg
     5761 agcatcaagt atatcgccgt gggcaaggcc gtgttccact ttgccatgga ggacctgggc
     5821 aagaccggcc aggatatcga gctgggcaag ctgagcaact ccgtgtctgg cggcctgacc
     5881 agcttcgact acgagcagat ccgggccgat gagacactgc agagacagct gtccgtggag
     5941 gtggcctttg ccgccaacaa tctgttcagg gcagtggtgg gacagaccgg caagaagatc
     6001 gagcagagca agtccgagga gaatgaggag gactttctgc tgtggaaggc cgagaagatc
     6061 gccgagtcca tcaagaagga gggcgagggc aacacactga agtctatcct gcagttcttt
     6121 ggcggcgcca gctcctggga tctgaatcac ttctgcgcag cctacggcaa cgagtctagc
     6181 gccctgggct atgagacaaa gtttgccgat gacctgagga aggccatcta ctccctgcgc
     6241 aatgagacat tccactttac cacactgaac aagggctctt ttgactggaa tgccaagctg
     6301 atcggcgata tgttcagcca cgaggcagca accggcatcg cagtggagag gacacgcttt
     6361 tacagcaaca atctgcctat gttctatcgg gagtccgacc tgaagagaat catggatcac
     6421 ctgtataaca cctatcaccc tcgcgccagc caggtgccat ctttcaacag cgtgtttgtg
     6481 aggaagaatt ttcgcctgtt cctgtccaac accctgaaca ccaatacatc tttcgacaca
     6541 gaggtgtacc agaagtggga gtccggcgtg tactatctgt ttaaggagat ctactacaac
     6601 tctttcctgc caagcggcga cgcccaccac ctgttctttg agggcctgag gcgcatcagg
     6661 aaggaggccg ataatctgcc catcgtgggc aaggaggcca agaagcgcaa cgccgtgcag
     6721 gactttggcc ggagatgcga tgagctgaag aacctgtctc tgagcgccat ctgtcagatg
     6781 atcatgaccg agtacaatga gcagaacaat ggcaacagga aggtgaagag cacacgggag
     6841 gacaagagaa agcccgatat cttccagcac tacaagatgc tgctgctgcg caccctgcag
     6901 gaggcctttg ccatctatat caggcgcgag gagttcaagt ttatcttcga cctgcccaag
     6961 acactgtacg tgatgaagcc cgtggaggag tttctgccta actggaagtc cggcatgttc
     7021 gactctctgg tggagcgggt gaagcagtcc cctgatctgc agagatggta tgtgctgtgc
     7081 aagttcctga atggccggct gctgaaccag ctgagcggcg tgatcagatc ctacatccag
     7141 tttgcaggcg acatccagcg gagagcaaag gcaaaccaca atcggctgta tatggataat
     7201 acccagagag tggagtacta ttctaacgtg ctggaggtgg tggacttctg catcaagggc
     7261 acatccaggt tctctaacgt gttcagcgat tacttccgcg atgaggacgc ctacgccgat
     7321 tatctggaca actatctgca gtttaaggac gagaagatcg ccgaggtgag cagcttcgcc
     7381 gccctgaaaa ccttctgtaa cgaggaggag gtgaaggccg gcatctacat ggacggcgag
     7441 aatcctgtga tgcagaggaa catcgtgatg gccaagctgt tcggccctga tgaggtgctg
     7501 aagaatgtgg tgccaaaggt gacacgggag gagatcgagg agtactatca gctggagaag
     7561 cagatcgccc catacagaca gaacggctat tgtaagtccg aggaggacca gaagaagctg
     7621 ctgaggttcc agcgcatcaa gaatcgcgtg gagtttcaga ccatcacaga gttctctgag
     7681 atcatcaacg agctgctggg ccagctgatc tcctggtctt ttctgcggga gagagatctg
     7741 ctgtactttc agctgggctt ccactatctg tgcctgcaca atgacaccga gaagcctgcc
     7801 gagtacaagg agatcagccg ggaggatggc acagtgatca gaaacgccat cctgcaccag
     7861 gtggcagcaa tgtacgtggg aggcctgcca gtgtataccc tggccgacaa gaagctggcc
     7921 gccttcgaga agggagaggc agactgtaag ctgagcatct ccaaggatac agccggcgcc
     7981 ggcaagaaga tcaaggattt ctttcggtac tccaagtatg tgctgatcaa ggacagaatg
     8041 ctgaccgatc agaaccagaa gtacacaatc tatctggccg gcctggagct gttcgagaat
     8101 accgatgagc acgacaacat cacagacgtg cggaagtacg tggatcactt taagtactat
     8161 gccacctctg acgagaatgc catgagcatc ctggatctgt attccgagat ccacgacaga
     8221 ttctttacat acgatatgaa gtaccagaag aacgtggcca atatgctgga gaacatcctg
     8281 ctgaggcact tcgtgctgat ccgccccgag ttctttaccg gcagcaagaa ggtcggcgag
     8341 ggcaagaaga tcacatgcaa ggccagggcc cagatcgaga tcgccgagaa tggcatgcgc
     8401 tccgaggact ttacctacaa gctgagcgat ggcaagaaga atatctccac atgtatgatc
     8461 gccgccaggg accagaagt
//
