LOCUS       sequence-189329-        7992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cg2Cas13a 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 7992)
  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 7992)
  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: pC025 - Cg2Cas13a from Carnobacterium gallinar...
FEATURES             Location/Qualifiers
     source          1..7992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(211..245)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        complement(join(3929..3934,3935..3952,3953..3958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      3996..4541
                     /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     4442..4461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4658..4675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5067..5169
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5170..5829
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5236..5255)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(join(6863..7992,1..61))
                     /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     7682..7701
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca
       61 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttgt
      121 tatagtcctc ttacatttag aggtagttta tattcgagac gaatctccgt ctccgttata
      181 gtcctcttac atttagaggt agtttatatt gctagcatta tacctaggac tgagctagct
      241 gtcaatcaat tggtaaacac cagcatcttc ttcaccagat cacagtactc ctgggacacc
      301 tggtccatag ggaaagagtg gttggacttc tccaggtgga tgatcttctt aggcttcagg
      361 ctctcaatct cgaagggctt tgtgtgatca ttgttggtca gcttcaggat catgccgtgc
      421 ttgtccagga tcttgatgat ggcctttgtc acggcattct tcagcttgcg atcgtaggac
      481 atcagcttcc tcaggtcgtt catgcagctc agcagagaca cgccctcgcc cttgccatca
      541 tttctcagca cgtgcaggtg ggcgatattg ttccggatct cgaagctgtg gttggtcacg
      601 cccagcaggg ccttcagctg gttctccatg ggcttcttca gattcttaaa ctcggctgtc
      661 tcctggaagg tcagatcgct agaggatgtg ccaaacaggt gcttattctc gttcacctcc
      721 cagtcgcgga tgttggcgaa ggtatcatag tcattcaggt agatagactg cttcttcagg
      781 gcgctataag atctgcccat atacacgaaa tcccgctcaa acagcacgga gaagcccacc
      841 atcctgccca tcaggtcgat catcagatag tgcaggttct tcaggtatgt caggcgcacc
      901 ttgtgatcca gcctattgaa gttctcgatg gcctcacaga tgatcttata ctcggcaaac
      961 ttggcgaact cggcgggctt gtccagcttc ttagacttgt tcttcttgat cttctcattg
     1021 tactcctcga tctctctggc cttggaggcc cactcctcgt gcagcttggt ccgcttatcc
     1081 accaggtcgg cgatcttctc cttctcctcg tgccagttca gcacgatctt ctcggacacc
     1141 ttaaactctg gattctcgat gaccaggttc tcgatgaaat tgcgtgtctg atacttctcc
     1201 agctgcttca taaacctgaa cttcacgggg gtcagcttgt cctcctgcat gtatggctca
     1261 ttcttggcca gctccacctt ctccagaaac acctcgatct tatcgaagga ctccttctcc
     1321 atggactcgc tgtcgttgct caggatgcac agctccagga ttggctgcat ctgctcgatc
     1381 agatcgccat tcttgatcag gcccttcttc acgctagact gcttaaactt gatcatctcg
     1441 tttctcagct cgctcaggtg ccttgcgtcc agcagctttg caaagatcca gaaggcgatc
     1501 tcctcatcgc cggtcttgat ctgctcgtcc acgttcagga tcttggcgat gcgctccttt
     1561 ctcttccgca ggccctcggc cttctccttc actgtgccca ccagctcctc ggcaggattc
     1621 aggttcagat cgaaattcct cagaaaggtg tcgaagccct tgataaagat ctggatcagc
     1681 agcttctcga aattgtagtt gggcttctcc tcgttggttt tcttggcctc cttgatggtg
     1741 cgctcctcct gcatctccct ctgcagatac tgcatgtact tctctgggga ctcgttcttc
     1801 atcagcttca cctgctcaaa ggcatacttt ctgatcttct tattgttctt cttatccttc
     1861 tctgtggtgt tcagctcctt attctgctgg atcacgtagt gcacggtgtc cttgaacagc
     1921 ttattgttgg cctggctaaa ctgtggcagg aacacgtagt ggtagatcag cttgatcaga
     1981 tagtagttgg ccttgaacac gtcattagag aaatcgtagt ctgtaaactg ctccagctcg
     2041 atggttttct cattctgata aaagtagccc tgcttataca gcttcttgta agaaggggcg
     2101 aaggccacca cctgtcctcc cagggacagt gtcagctcct tgttgctgaa cagcttctcc
     2161 aggtccttca tggaatactg ctccagggcg ccggagctct tgatcttctc ggcaaagtag
     2221 atctcgatat ccttgatctc ggcgtcgaac agcttctgat agatggtttt ctggtaagac
     2281 acgttgccca gatcgccatt ctcaaaggag ccgaccttca gctggttggg ctcgtgtctc
     2341 ttattgatct gctggtggaa gatctgattg cgcaccctct gcacggctcc cctgattgcc
     2401 cacaggtcca gctcgtttgg tgtcatggcg atcttcttgg caggattctc atcgcgctcc
     2461 tccacgatct cggtcttaaa gaacaggtcg tgcttcagct ccacgttctt ctccagctcc
     2521 ttcttaaact tctctctggt caggatgtcc tcgtttgtgg tggcctggat gatattccgc
     2581 agagaggagc tggcgaacac gcaggcatcc ataaactttg tctggaagcc ctcatacatg
     2641 ccgatctcct gcagggtctg gctgtccagc acgcccttgt tgtactggtg cagcttgccc
     2701 acttgcagca gatactgcac gaaggcgttc ctcagctggt agcccactgt cttgatgatg
     2761 gtctgtggat tcagatagta ggactcggcg ctctccttct tgcctgtctt aggaaaatag
     2821 tgctcgaagt acttgcccag ctcgatgtta aactgattca gggcctcgtt gccctcctga
     2881 ttgaacagct tgatcagggc cttatccttg ttgtgagggt gctgcttcca cagcttcttg
     2941 atctcaaagt ttctggtctt attggactta tccttgtcga agttcatcag cttctctgtg
     3001 atggcggtgc acagctcgtc cacgttatac acggcggcca gctcgttgat cttattggca
     3061 aactccttat tgaactgctc cacggcaaaa ttgttctgct ggatcagagg gctctcctga
     3121 aacttcatgg cccactgctg gaacagctgc ttccgcttag acagcaggtt aggctcctct
     3181 gcggttggca tgctgaagcc cctgttctgc tggatagaat tggccaggat ctccttcttc
     3241 ctattcagat agctcttctt ataatccagg tagggagaca tctccttgca aaagtaggac
     3301 cagttcttct catcgaactt gtcctcctgg attctgtcca cggcgttaaa gaacaggggg
     3361 atcagattca gctttgttgg cacgctctga ttcttctcag aggtatactt gatctcctcg
     3421 tggtacttgc tcttaaagaa ccgcttcagg ccggcctcct tcagtctccg gatattgttc
     3481 aggtacacgc tgggcttctc agagttggac ttctccagct gctcaaagat gatgaagata
     3541 ttctcggcca gcaccttgta gcccttcacc tggtcggcct tcaccagggt cttgttcaga
     3601 aaggaggcgg cgaagctctg ctccttcttt gtcaggatgt tcacggtatt tgtggtgcca
     3661 ttccacacca ggtggccgtt cagcttgctg cgattcatgc tcacggggct gccgttgatc
     3721 ttcacctttg tcatcctcat gttcagctga ttgccggggt ttgtggtcag cagggccagc
     3781 accagctggc agtactcctc ggacacctgg ttggtcttga tggccttatc cttcaggtgg
     3841 atgatctcct ttgggatcag gctctcgatc tcgaaattct tgtggttctc gtcgatcttc
     3901 agcttcagga tcatggataa gacctcctaa catacgagcc ggaagcataa agtgtaaatc
     3961 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     4021 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     4081 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     4141 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     4201 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     4321 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     4381 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     4441 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     4501 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     4561 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     4621 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     4681 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     4741 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     4801 gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca gtagctgaac
     4861 aggagggaca gctgatagaa acagaagcca ctggagcacc tcaaaaacac catcatacac
     4921 taaatcagta agttggcagc atcacccgac gcactttgcg ccgaataaat acctgtgacg
     4981 gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg
     5041 gccaactttt ggcgaaaatg acacgttgat cggcacgtaa gaggttccaa ctttcaccat
     5101 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     5161 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     5221 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     5281 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     5341 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     5401 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     5461 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     5521 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     5581 tttattgaga atatgttttt cgtgtcagcc aatccctggg tgagtttcac cagttttgat
     5641 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     5701 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     5761 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     5821 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg ctacgcctga
     5881 ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg
     5941 gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga
     6001 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc
     6061 tccccgtgga ggtaataatt gacgatatga tcatttattc tgcctcccag agcctgataa
     6121 aaacggttag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     6181 tgcgatgcag atccggaaca taatggtgca gggcgcttgt ttcggcgtgg gtatggtggc
     6241 aggccccgtg gccgggggac tgttgggcgc tgccggcacc tgtcctacga gttgcatgat
     6301 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     6361 accggcagcg gtgcggactg ttgtaactca gaataagaaa tgaggccgct catggcgttg
     6421 actctcagtc atagtatcgt ggtatcaccg gttggttcca ctctctgttg cgggcaactt
     6481 cagcagcacg taggggactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     6541 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     6601 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     6661 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc
     6721 cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc
     6781 gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta
     6841 gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca
     6901 acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg
     6961 tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc
     7021 tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg
     7081 acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg
     7141 ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg
     7201 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     7261 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     7321 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     7381 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     7441 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgacgc tctcccttat
     7501 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     7561 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     7621 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     7681 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     7741 cgatgcgtcc ggcgtagagg atccacagga cgggtgtggt cgccatgatc gcgtagtcga
     7801 tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg
     7861 ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat
     7921 agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca
     7981 taaccaagcc ta
//
