LOCUS       sequence-181301-        8181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RcsCas13a 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: pC030 - RcsCas13a from Rhodobacter capsulatus ...
FEATURES             Location/Qualifiers
     source          1..8181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(122..156)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        complement(join(4029..4034,4035..4052,4053..4058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      4096..4641
                     /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     4542..4561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4758..4775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5167..5269
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5270..5929
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5336..5355)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(6963..8153)
                     /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     7782..7801
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aactgtgata aaataccgca ttaaagcttg ttcagtccgc cgtcgtcttg gcggtgatgt
       61 gaggccgaga cgaatctccg tctccgttca gtccgccgtc gtcttggcgg tgatgtgagg
      121 cgctagcatt atacctagga ctgagctagc tgtcaatcag tcaattcgca ccaggtcagc
      181 ggcgttcagt ttggaagtgt tgctcttggg ggggacatag atttcgaccc tgaatctgta
      241 tctccgcccg acaatgatgg cgtcctctct gattcgtcgc aggcgggaga tatgcaggcc
      301 aatattgcca gccaccccct ccacggccag aaagcccagt gagccggtat caatgacttt
      361 gaccacgact ccttcgtaga cctccccgac ctgtgggggt ggcagcctgg accctgcaga
      421 aggtgctttg ttgggaggtt tctgatcagg ctgagactga gcagttgcag gcttgggtgg
      481 cttgtgccag gcgtccccat catcccggcg ccgtggttta gggttctgca cgctcccatt
      541 gaacactgca gcgaccatct gcaggtagtc ctgtgagaag cgtgcttcgg tgacggcggc
      601 ggggcccccc acggtcagcc gcactttatc caggtgtttg attgcagcct gggtgattgt
      661 agcgtcctgc agcaggtggt ccttcatctg ccacttcagt gtcagcccca gccgagccag
      721 catatccagg atactccgtg ggactgcgtt tttgcgtttg cgatcgtagg ccatcatttc
      781 tcttgccctg ttcaccagag ctgtcaggtc gggggttccg tcggctctat ccagcacgtt
      841 gaaatgtgcc aggtctttcc gggtctgggt gcgtgcgtcg gtattagcgg cgcctgcaat
      901 ggtcactgcc caatcagcgc cagcccccag ctgtttgctg gcgttgatca ggaaagtgcc
      961 agtatcgcgc tcataagcgc cagcgtaatc aatcaggcgg ccgatgacat ccatcatcag
     1021 gcgatgcagc cgcaggtgat cccccaggta cactctgccc accaggaacc tgtgctcctc
     1081 aatggttttg attgcggcgg cgtaactctg ccgctcctcg gggctgatgg ttttagggtg
     1141 gcatttcatg actttatcgt gcagatctgt cctcagttcc tgtgcggcga caatctgaga
     1201 tttttcctga gtttcgtcct caatctcggc caggcgtgcg acttcctcat cccgcacttt
     1261 atgtttagcg aacagatcgc tcagcactgc catgtggtta tatcgtgcaa tctggcgcag
     1321 cccgcgcagt gtcctggcga cccgcagctc tggttcggag gctccgtcgc cctcggggtc
     1381 atcttcagcg ggcctagcag tggttggtgt ggccataaac agccgatcga aagtagctgg
     1441 attagcaaac agtgcccgga aaggcttcag atcaaatgga gcggcgcggc cttcaaatct
     1501 ggcttcccct gttttcagaa acaggaccag cacatcgcgg aaccttttca ccagatccag
     1561 ggcctccctc cgggcatctg cctggtctgt ggcgtcccca tcctggacca gctcgtattt
     1621 gccctgcagt gcttcccact ttctcagctg gtgcagcagg ttgctcacat cactagctgg
     1681 gacgaagtgc atgaccaggt acagtgcggc ctgccagtgt tcatactggc ctcgtgtatc
     1741 gattggctct ggcagcacag gggctctggt cattgctgtg gccccaggtt caatcttcag
     1801 aatccagtcg aacccctttg ctcgaatgta atcctcgaac atgaaggcca gcatatctcg
     1861 cctatagttc tcgataaact cggcctgctt tctagcattc tctgagtcgg actcatagcc
     1921 aatctgcacc cggaactctg tggcagtctc gccagtcagt gcactcaggt attccctcag
     1981 ggtttcgcca tcattaggtg gcctcagtcg tgaggtgcgt ccctccatca catcggaata
     2041 tgctttagtg acattcacgg actgggccag tgcagtagct gcctctttag ctctggcagc
     2101 aggtcctgcc agggcagttc cagtgattcc acttgcgtag gcccgaaatg ggccatcata
     2161 cagccgcagc agagcgatat acctggcctt tgttgcaggg tcgtccagtt cccgtggagc
     2221 tggaggaggt ggcagcttag tggcgaaagc atgttttcgg gtatcccgca gtccatgcac
     2281 gtatcccctg actccgtctg cccgtttcag cagcagcttc agccgaggca gcccggatga
     2341 cacttcagga gcgtctttca ctgctttgac gatctcggaa tacagtgtgc tgaagtgttc
     2401 tttgcttgca tagtgtgcca caaaagctcc gctcaggtct gccagcagcc gggcgcccag
     2461 ggctttggcg tcattgtcaa tgattgctct aatggctgcc actgtcttgt cggtcaggac
     2521 cacatgctcg gcttcttttg ccttgcccca ccgtgtcttt tccagctcct tgcggatttc
     2581 cttcagaaat cctgcccgag cccccagatg aaaagtctgg tttctacacc ctctcaggta
     2641 tcgcagcaga gcgaacacga agccttcatt tccctctgcc cccagtctat ccagctctgg
     2701 tgtctcgccg aaatagattc cgcgccgagc cattgcctca gcgaccatct ccttgttgga
     2761 gattacctgt ctgatgttga cggcggcagt cagatctcta tcattcttct cagtgttcac
     2821 gatcttcccc attgggtcaa tccatgcttt catgctgcgc ccagccagtg caaaagctcc
     2881 cacccacagc ctgacaaaga tctcgctttc tttgatttct gtctggccgg cacttgtcca
     2941 atagtgggac tgagccagat ccccggcctg ctggcctcgg tattctgaca cccttcccat
     3001 tctgaccatc tgattccgga ctctgttctc atgggtatgt ctcatcagag ccagcagttc
     3061 ggtcttgtcg gctggaaaag ctctggctgc attctttccc cgggcgcaca gtcttttata
     3121 agttttcttc acttcgtcgt gcagtgccag gacagaaggg ccggctgcgg gatcgaatga
     3181 gcacctcttg gagcctgtca ccctttccag atgctctgcc agtgcgaacc ccactttggg
     3241 actcagactt ctcccgctga cctggcgagg tggatcgaaa gcctgtgctg cgctcacagc
     3301 agctttagcc acctccttca ctgatgcggc cacatccacg acaaagtaag tctggatttc
     3361 ctcttcgccc caattcctag ccagtctagc cattcctctt ggcagcacgc ttgactcaat
     3421 ccccagagcg cgggccacca ccagccctgg agcgaattta tcagtcttag gattccgctt
     3481 aggctggcca tcgattcttt tctcatcgac gtgcagatgt tcgtacagag cgactgccag
     3541 ctcctttgcg tcattcccgc gcttggacat tgctccatac cacctgccgt ggaaggcctg
     3601 aggctcaggt gggtcggctt cgaacttcag ttttgctccg ctatcagcag gctgaaattt
     3661 gtctccgtat ggatgcagtc gccgcttaaa ctgatcatag tcactatcct gagccaggcc
     3721 agcttctgac accagtttcc agaatgcctc tcccaggtca tcccttgcgt caaactgcat
     3781 tttgctaggg gttggggagt ggattgcttt gccatcggat ttcctactat cgggcttccg
     3841 gtagattttg tcgatccctg agatccactg tccaatcagg gcctttggat cacttgacag
     3901 gtgtgctggc agttccttgc gattctttcc atcggtgcta atctttctct tcagtcctcc
     3961 tgcggggtcc ccaaactcgc taatagtcct tccctgcact tttccaatct gcatggataa
     4021 gacctcctaa catacgagcc ggaagcataa agtgtaaatc tagaaatatt ttatctgatt
     4081 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
     4141 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg
     4201 aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc
     4261 ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg
     4321 cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4381 cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac
     4441 tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac
     4501 cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt
     4561 tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca
     4621 ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta
     4681 tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca
     4741 gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca
     4801 tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac
     4861 accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggcg
     4921 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa
     4981 acagaagcca ctggagcacc tcaaaaacac catcatacac taaatcagta agttggcagc
     5041 atcacccgac gcactttgcg ccgaataaat acctgtgacg gaagatcact tcgcagaata
     5101 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg
     5161 acacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     5221 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     5281 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     5341 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     5401 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     5461 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
     5521 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     5581 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     5641 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     5701 cgtgtcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     5761 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     5821 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     5881 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     5941 cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat
     6001 gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa
     6061 atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc
     6121 gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt
     6181 gacgatatga tcatttattc tgcctcccag agcctgataa aaacggttag cgcttcgtta
     6241 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     6301 taatggtgca gggcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     6361 tgttgggcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     6421 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct accggcagcg gtgcggactg
     6481 ttgtaactca gaataagaaa tgaggccgct catggcgttg actctcagtc atagtatcgt
     6541 ggtatcaccg gttggttcca ctctctgttg cgggcaactt cagcagcacg taggggactt
     6601 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     6661 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     6721 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     6781 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat
     6841 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     6901 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     6961 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     7021 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     7081 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     7141 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     7201 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     7261 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     7321 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     7381 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     7441 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     7501 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     7561 ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg cattaggaag
     7621 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     7681 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     7741 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     7801 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     7861 atccacagga cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa
     7921 gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat
     7981 agaaattgca tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg
     8041 gaatggacga tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca
     8101 gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt
     8161 gcctgactgc gttagcaatt t
//
