LOCUS       sequence-181260-        8181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RcdCas13a 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 8181)
  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 8181)
  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: pC032 - RcdCas13a from Rhodobacter capsulatus ...
FEATURES             Location/Qualifiers
     source          1..8181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1282..1287,1288..1305,1306..1311))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      1349..1894
                     /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     1795..1814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2011..2028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2420..2522
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2523..3182
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2589..2608)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(4216..5406)
                     /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     5035..5054
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(5556..5590)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 tttccttatt gctgatgacc tgcctgatgt tcactgctgc agtcaggtcc cggtcgttct
       61 tctcagtgtt cacgatcttc cccatagggt cgatccaggc cttcatggat cggcctgcca
      121 gggcaaatgc cccgacccac agccgcacga agatttcgct ctccttaatc tcggtctgcc
      181 ctgcgctagt ccaataatgg ctctgagcca gatccccggc ctgctgtccc cgatactcgc
      241 tgaccctccc catcctgacc atctggttgc gcaccctatt ttcgtgtgtg tgtctcatca
      301 gtgccagcag ctctgtcttg tctgctggaa aagctcttgc ggcatttttc cctctggcgc
      361 acagtcgctt ataggtcttt ttcacctcgt cgtgcagtgc caggacactt ggcccagctg
      421 ctggatcaaa gctgcatctt ttgcttccgg tgaccctttc caggtgctct gccagggcaa
      481 acccgacctt tggagacagg ctccgtcctg acacctgtcg aggtgggtca aatgcctgag
      541 cagcggacac ggcagccttg gccacttcct tcactgaggc tgccacgtcc acgacgaaat
      601 aagtctgaat ttcctcctct ccccagttcc tggccagcct ggccatcccc cttggcagca
      661 ctgagctctc gatccccagt gcccttgcga ccaccagccc gggggcaaat ttatcagtct
      721 ttgggttcct ctttggctgg ccatcaatcc ttttttcgtc cacatgcaga tgctcataca
      781 gtgcagctgc cagttccttt gcgtcgttcc ccctctttga catggcccca taccagcgtc
      841 cgtggaaagc ctgtggctca gggggatcgg cctcgaactt cagttttgcg ccgctgtcag
      901 cgggctgaaa cttgtcccca taagggtgca gcctgcgctt aaactgatca taatctgagt
      961 cctgtgccag ccctgcttca ctcaccagtt tccagaatgc ttcccccaga tcatcccggg
     1021 cgtcgaactg cattttgctt ggtgtaggac tgtggatagc cttgccgtct gattttctgc
     1081 tatctggttt ccggtagatc ttgtcaatgc cactaatcca ctggccaatc agagcctttg
     1141 gatcgcttga caggtgagct ggcagttctt tccggttctt tccgtcagta gaaatcttcc
     1201 tcttcagtcc tcctgctggg tccccgaact cgctaatagt ccttccctgc actttgccga
     1261 tctgcatgga taagacctcc taacatacga gccggaagca taaagtgtaa atctagaaat
     1321 attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     1381 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     1441 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     1501 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     1561 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     1621 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     1681 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     1741 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     1801 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     1861 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     1921 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     1981 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     2041 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     2101 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg
     2161 ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg
     2221 acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca
     2281 gtaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc
     2341 acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact
     2401 tttggcgaaa atgacacgtt gatcggcacg taagaggttc caactttcac cataatgaaa
     2461 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     2521 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     2581 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     2641 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2701 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     2761 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     2821 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     2881 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2941 agaatatgtt tttcgtgtca gccaatccct gggtgagttt caccagtttt gatttaaacg
     3001 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     3061 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     3121 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     3181 aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg
     3241 ataataagcg gatgaatggc agaaattcga aagcaaattc gacccggtcg tcggttcagg
     3301 gcagggtcgt taaatagccg cttatgtcta ttgctggttt accggtttat tgactaccgg
     3361 aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag tttgctcagg ctctccccgt
     3421 ggaggtaata attgacgata tgatcattta ttctgcctcc cagagcctga taaaaacggt
     3481 tagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     3541 cagatccgga acataatggt gcagggcgct tgtttcggcg tgggtatggt ggcaggcccc
     3601 gtggccgggg gactgttggg cgctgccggc acctgtccta cgagttgcat gataaagaag
     3661 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctaccggca
     3721 gcggtgcgga ctgttgtaac tcagaataag aaatgaggcc gctcatggcg ttgactctca
     3781 gtcatagtat cgtggtatca ccggttggtt ccactctctg ttgcgggcaa cttcagcagc
     3841 acgtagggga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     3901 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     3961 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4021 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc
     4081 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca
     4141 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt
     4201 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
     4261 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc
     4321 cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag
     4381 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
     4441 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
     4501 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc
     4561 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     4621 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     4681 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     4741 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4801 ggagctgact gggttgaagg ctctcaaggg catcggtcga cgctctccct tatgcgactc
     4861 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     4921 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     4981 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     5041 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     5101 tccggcgtag aggatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc
     5161 tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag
     5221 aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact
     5281 ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa
     5341 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     5401 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     5461 cttgttcagt ccgccgtcgt cttggcggtg atgtgaggcc gagacgaatc tccgtctccg
     5521 ttcagtccgc cgtcgtcttg gcggtgatgt gaggcgctag cattatacct aggactgagc
     5581 tagctgtcaa tcaatcaatc cgcaccaggt cggcagcgtt cagtttggaa gtgttgctct
     5641 ttggaggcac gtagatttcc acccggaacc tataccttcg tccgacaatg atagcatcct
     5701 cccgaatccg cctcagtcgg gagatgtgca ggccaatatt tccagcgacg ccttcgacag
     5761 ccaggaaccc cagagatcca gtatcaatga ctttgacgac cacgccttcg tagacctctc
     5821 ccacctgtgg agggggcagc cgggaccctg cggaaggtgc tttgttggga ggtttctgat
     5881 ctggctggct ctgggcggtg gctggcttag gaggtttatg ccaagcatct ccgtcgtccc
     5941 tgcgccgtgg cttggggttc tgcactgatc cgttaaacac tgcagccacc atctgcaggt
     6001 agtcctgaga gaaccgagct tctgtgactg ctgctggtcc gcccacggtc agtctcacct
     6061 tgtccaggtg cttgatagct gcctgggtaa tggttgcatc ctgcagcagg tgatccttca
     6121 tctgccactt cagggtcagt cccagccgtg ccagcatatc cagaatgctc cttgggacgg
     6181 cattttttct cttccgatcg taggccatca tttcccgtgc gcgattcacc agtgcagtca
     6241 ggtcgggggt cccatcagcc cgatccagca cgttgaagtg agccagatcc ttccgtgtct
     6301 gagttctagc atctgtattg gcggccccgg cgattgtcac tgcccagtct gcgccggccc
     6361 ccagctgttt gcttgcattg atcaggaatg tgccagtatc tctttcatag gctccagcat
     6421 agtcaatcag gcggccgatc acgtccatca tcagccggtg cagtcgcaga tgatccccca
     6481 gatagaccct ccccaccaga aatctatgct cttcgatagt tttgatggct gctgcgtagc
     6541 tctggcgctc ctcggggcta attgtcttag ggtggcactt catgaccttg tcgtgcaggt
     6601 cggtccgcag ctcctgtgcg gcgacgatct gtgacttctc ctgggtttca tcctcaatct
     6661 cggccagcct tgccacctct tcatcgcgca ccttatgttt ggcgaacagg tctgacagga
     6721 ctgccatgtg attgtaccgt gcgatctgcc gcagcccccg cagtgtcctt gcgacccgca
     6781 gttcaggctc ggatgccccg tccccctctg gatcatcctc ggctggtcga gctgtggtag
     6841 gtgttgccat aaacaggcga tcgaatgttg cggggttggc aaacagagct cggaatggct
     6901 tcaggtcaaa aggagcagcc cttccctcga agcgtgcctc gccagttttc aggaacagca
     6961 ccaggacatc tcggaaccgc ttgaccaggt ccagagcttc ccgccgggcg tcggcctggt
     7021 ctgtggcgtc cccgtcctgg accagctcgt acttcccctg cagggcttcc catttccgca
     7081 gctggtgcag caggttagac acgtctgagg ctggcacgaa atgcatgacc agatacagtg
     7141 cagcctgcca atgctcgtac tgccctcgtg tatcgatagg ttcgggcagc actggtgccc
     7201 tagtcatggc tgttgccccg ggctcaattt tcagaatcca atcgaacccc ttggctctga
     7261 tataatcctc gaacataaaa gccagcatat ctctccggta attctcaata aactcagcct
     7321 gttttcttgc gttttcgctg tcgctttcgt atccgatctg gacccgaaac tctgttgcag
     7381 tttccccagt cagtgcagac aggtattctc tcagagtctc tccgtcattg ggaggcctca
     7441 gccgtgatga ccgtccttcc atcacgtcag agtaggcctt agtgacattc acggactgag
     7501 ccagagctgt tgctgcctct ttggccctag ctgctgggcc agccagtgca gtccctgtaa
     7561 tcccggatgc gtaagctctg aatgggccgt catacaggcg cagcagggcg atgtacctgg
     7621 ccttggttgc gggatcgtcc agctctcttg gggcaggtgg gggtggcagc tttgttgcga
     7681 aggcatgttt tcttgtatcc ctcagcccgt ggacataccc gcgcacccca tcggcccttt
     7741 tcagcagcag cttcagcctg ggcagtcctg aactgacttc aggtgcatcc ttcacagcct
     7801 tgacgatctc agaatacagt gtagaaaagt gctccttgct tgcgtagtgg gcgacaaagg
     7861 cgccggacag atctgccagc agtctggccc ccagtgcttt ggcatcattg tcgatgatag
     7921 ctcggatagc tgccacagtt ttgtctgtca ggaccacatg ctctgcctct ttggccttcc
     7981 cccaccgggt cttttccagc tctttccgaa tctctttcag aaagccagct cttgccccca
     8041 gatgaaatgt ctgatttcta cagcccctca ggtatcgcag cagagcgaac acgaacccct
     8101 cgtttccttc tgcccccagt cggtccagct ctggagtctc cccgaagtag attccccttc
     8161 tagccattgc ctcggcgacc a
//
