LOCUS       sequence-181268-        8635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LbnCas13a 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 8635)
  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 8635)
  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: pC022 - LbnCas13a from Lachnospiraceae bacteri...
FEATURES             Location/Qualifiers
     source          1..8635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3348..3382
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             3530..4720
                     /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(3882..3901)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5754..6413)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6328..6347
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6414..6516)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6908..6925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7042..7587)
                     /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(7122..7141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7625..7630,7631..7648,7649..7654)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 acgtgtggga ggaccacgca gcccggagag tggacaatag ggagttcatc aagctgccac
       61 tggagaacaa gctggccaat ggcaagaccg acaaggatgc cgagcggatc agaaagaaca
      121 cagtgaagga gctgtataga aaccagaata tcggctgcta caggcaggcc gtgaaggccg
      181 tggaggagga caacaatggc cggtactttg acgataagat gctgaacatg ttctttatcc
      241 acagaatcga gtatggcgtg gagaagatct acgccaatct gaagcaggtg accgagttca
      301 aggcccgcac aggctacctg agcgagaaga tctggaagga tctgatcaac tacatctcta
      361 tcaagtatat cgccatgggc aaggccgtgt ataactatgc catggatgag ctgaatgcct
      421 ccgacaagaa ggagatcgag ctgggcaaga tctccgagga gtatctgtct ggcatctcta
      481 gcttcgacta cgagctgatc aaggccgagg agatgctgca gagggagaca gccgtgtacg
      541 tggcctttgc cgcaaggcac ctgtcctctc agacagtgga gctggattcc gagaactctg
      601 acttcctgct gctgaagcct aagggcacca tggacaagaa cgataagaat aagctggcct
      661 ccaacaatat cctgaatttt ctgaaggata aggagacact gcgggacaca atcctgcagt
      721 atttcggcgg ccactctctg tggacagatt tcccatttga caagtacctg gccggcggca
      781 aggacgatgt ggattttctg accgacctga aggatgtgat ctatagcatg cggaacgact
      841 ccttccacta cgccacagag aaccacaaca atggcaagtg gaataaggag ctgatctccg
      901 ccatgtttga gcacgagaca gagagaatga cagtggtcat gaaggacaag ttctattcta
      961 acaatctgcc catgttttac aagaacgacg atctgaagaa gctgctgatc gacctgtata
     1021 aggacaatgt ggagagagcc tctcaggtgc ccagcttcaa caaggtgttt gtgcgcaaga
     1081 atttccctgc cctggtgcgg gacaaggata acctgggcat cgagctggat ctgaaggccg
     1141 acgccgataa gggcgagaat gagctgaagt tctacaacgc cctgtactac atgttcaagg
     1201 agatctacta caacgccttc ctgaacgaca agaatgtgag ggagcggttc atcaccaagg
     1261 ccacaaaggt ggccgacaac tatgatagga ataaggagcg caacctgaag gataggatca
     1321 agagcgccgg ctccgacgag aagaagaagc tgcgcgagca gctgcagaat tacatcgccg
     1381 agaacgattt cggccagagg atcaagaata tcgtgcaggt gaaccctgac tataccctgg
     1441 cccagatctg ccagctgatc atgacagagt acaaccagca gaacaatggc tgtatgcaga
     1501 agaagagcgc cgcccggaag gatatcaata aggactccta ccagcactat aagatgctgc
     1561 tgctggtgaa cctgagaaag gccttcctgg agtttatcaa ggagaattat gcctttgtgc
     1621 tgaagcccta caagcacgac ctgtgcgata aggccgactt cgtgcctgat tttgccaagt
     1681 acgtgaagcc ctacgccggc ctgatcagca gggtggcagg cagctccgag ctgcagaagt
     1741 ggtatatcgt gtcccgcttt ctgtctcctg cccaggccaa ccacatgctg ggcttcctgc
     1801 actcctacaa gcagtacgtg tgggacatct acaggcgcgc ctctgagaca ggcaccgaga
     1861 tcaatcacag catcgccgag gataagatcg ccggcgtgga catcaccgac gtggatgccg
     1921 tgatcgatct gagcgtgaag ctgtgcggca caatctctag cgagatctcc gactacttca
     1981 aggacgatga ggtgtacgcc gagtatatct cctcttacct ggattttgag tatgacggcg
     2041 gcaactacaa ggatagcctg aataggttct gtaactccga tgccgtgaat gaccagaagg
     2101 tggccctgta ctatgacggc gagcacccta agctgaaccg caatatcatc ctgagcaagc
     2161 tgtacggcga gcggagattt ctggagaaga tcaccgatcg ggtgagccgg agcgacatcg
     2221 tggagtacta taagctgaag aaggagacaa gccagtacca gacaaagggc atcttcgata
     2281 gcgaggacga gcagaagaac atcaagaagt ttcaggagat gaagaatatc gtggagttca
     2341 gagatctgat ggactatagc gagatcgccg atgagctgca gggccagctg atcaactgga
     2401 tctacctgcg ggagcgggac ctgatgaatt tccagctggg ctaccactat gcctgcctga
     2461 acaatgattc caacaagcag gccacctatg tgacactgga ctaccagggc aagaagaatc
     2521 ggaagatcaa cggcgccatc ctgtatcaga tctgtgccat gtatatcaat ggcctgcctc
     2581 tgtactatgt ggacaaggat agctccgagt ggaccgtgtc tgacggcaag gagagcacag
     2641 gcgccaagat cggcgagttc tacagatatg ccaagtcctt tgagaacacc tctgattgct
     2701 acgccagcgg cctggagatc tttgagaata tcagcgagca cgacaacatc acagagctga
     2761 ggaattatat cgagcacttt cgctactaca gcagcttcga ccggagcttc ctgggcatct
     2821 actctgaggt gttcgatcgg ttctttacct acgacctgaa gtatagaaag aacgtgccaa
     2881 caatcctgta taacatcctg ctgcagcact tcgtgaacgt gaggttcgag ttcgtgagcg
     2941 gcaagaagat gatcggcatc gataagaagg accgcaagat cgccaaggag aaggagtgtg
     3001 cccggatcac catcagagag aagaacggcg tgtatagcga gcagtttacc tacaagctga
     3061 agaatggcac agtgtatgtg gatgccaggg acaagagata cctgcagtcc atcatcaggc
     3121 tgctgttcta tccagagaag gtgaacatgg atgagatgat cgaggtgaag gagaagaaga
     3181 agccctccga caacaatacc ggcaagggct actctaagag ggatcgccag caggaccgca
     3241 aggagtacga taagtataag gagaagaaga agaaggaggg caatttcctg agcggcatgg
     3301 gcggcaacat caattgggac gagatcaatg cccagctgaa gaactgattg acagctagct
     3361 cagtcctagg tataatgcta gcgttgatga gaagagccca agatagaggg caataacgga
     3421 gacggagatt cgtctcggtt gatgagaaga gcccaagata gagggcaata acaagcttta
     3481 atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa
     3541 caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt
     3601 tatgccggta ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca
     3661 ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg
     3721 agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac
     3781 tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atcctctacg ccggacgcat
     3841 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     3901 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     3961 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     4021 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     4081 gcataaggga gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     4141 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     4201 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     4261 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     4321 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     4381 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     4441 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     4501 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     4561 cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     4621 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     4681 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     4741 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     4801 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     4861 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     4921 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     4981 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     5041 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcccc tacgtgctgc
     5101 tgaagttgcc cgcaacagag agtggaacca accggtgata ccacgatact atgactgaga
     5161 gtcaacgcca tgagcggcct catttcttat tctgagttac aacagtccgc accgctgccg
     5221 gtagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     5281 ttatcatgca actcgtagga caggtgccgg cagcgcccaa cagtcccccg gccacggggc
     5341 ctgccaccat acccacgccg aaacaagcgc cctgcaccat tatgttccgg atctgcatcg
     5401 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctaaccgt
     5461 ttttatcagg ctctgggagg cagaataaat gatcatatcg tcaattatta cctccacggg
     5521 gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     5581 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     5641 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt
     5701 attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc
     5761 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     5821 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     5881 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     5941 aaatcaaaac tggtgaaact cacccaggga ttggctgaca cgaaaaacat attctcaata
     6001 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     6061 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     6121 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     6181 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     6241 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     6301 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     6361 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     6421 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     6481 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt gtcattttcg ccaaaagttg
     6541 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     6601 tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt
     6661 tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc
     6721 ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta
     6781 gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat
     6841 gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt
     6901 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     6961 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     7021 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     7081 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7141 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     7201 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     7261 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     7321 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     7381 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     7441 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     7501 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     7561 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     7621 tagatttaca ctttatgctt ccggctcgta tgttaggagg tcttatccat gaagatcagc
     7681 aaggtgcggg aggagaacag aggcgccaag ctgaccgtga atgccaagac agccgtggtg
     7741 tctgagaacc ggagccagga gggcatcctg tataatgacc caagcaggta cggcaagtcc
     7801 cgcaagaacg acgaggatag ggaccgctat atcgagagca gactgaagag ctccggcaag
     7861 ctgtaccgga tctttaacga ggataagaat aagcgcgaga cagacgagct gcagtggttc
     7921 ctgtccgaga tcgtgaagaa gatcaaccgg agaaatggcc tggtgctgtc cgacatgctg
     7981 tctgtggacg atagagcctt cgagaaggcc tttgagaagt acgccgagct gtcttatacc
     8041 aacaggcgca ataaggtgtc cggctctccc gccttcgaga catgcggagt ggatgcagca
     8101 acagcagagc ggctgaaggg aatcatctcc gagacaaact tcatcaacag aatcaagaac
     8161 aacatcgaca acaaggtgtc tgaggacatc atcgatcgga tcatcgccaa gtacctgaag
     8221 aagagcctgt gccgggagag agtgaagaga ggcctgaaga agctgctgat gaatgccttt
     8281 gatctgccat actctgatcc cgacatcgat gtgcagaggg acttcatcga ttatgtgctg
     8341 gaggactttt accacgtgcg ggccaagagc caggtgagca gatccatcaa gaacatgaat
     8401 atgcccgtgc agcctgaggg cgacggcaag ttcgccatca ccgtgagcaa gggcggcaca
     8461 gagtctggca acaagcgcag cgccgagaag gaggccttca agaagtttct gagcgattac
     8521 gcctccctgg acgagagggt gcgcgacgat atgctgcgga gaatgaggcg cctggtggtg
     8581 ctgtactttt atggctccga cgattctaag ctgagcgatg tgaacgagaa gttcg
//
