LOCUS       sequence-189334-        8477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LshCas13a 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 8477)
  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 8477)
  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: pC018 - LshCas13a from Leptotrichia shahii
FEATURES             Location/Qualifiers
     source          1..8477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(196..230)
                     /label=BBa_J23111
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.58"
     promoter        complement(196..230)
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     promoter        complement(196..230)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        complement(join(4415..4420,4421..4438,4439..4444))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      4482..5027
                     /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     4928..4947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5144..5161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5553..5655
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5656..6315
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5722..5741)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(join(7349..8477,1..62))
                     /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     8168..8187
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
       61 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagcttg
      121 ttttagtccc cttcgatatt ggggtggcga gacgaatctc cgtctccgtt ttagtcccct
      181 tcgatattgg ggtgggctag cactatacct aggactgagc tagctgtcaa tcacagggta
      241 tcgttagtat tctcgatttt ggtcagcagt tcaatgatca gattcttaat gtagtcactg
      301 ttgtaggact ccagttccag gacgctcact ttcttgggct tcatcaggcg ctccagaata
      361 tcattgttgc cgatcagttt gaatttcttt ttcagctcat cgtagtccag attcacgtcc
      421 tttttaaaga cttcgaacac gctggcataa gttgaattgt tgtagcgtgt gctatatgac
      481 agcaggttgc tcacccgatc gatctgctca gcaatactgt agtcggcaaa tggatttctc
      541 acgatgtaga aatgggagat atagttccga atgctctcat tttcaggctt gttgatctcg
      601 gaattttcgc tcaggtcgat tccaaacccg tagcaaatct tctcgaactt cttgtaggac
      661 tcttcatcaa agaacttgta gtaggctgtg gtagtgtaaa agccgtcgct tccattgcgc
      721 tttgggtatg ccctgctgat cccggtgtta tagcctgaca gcttaatgat gcccagctcc
      781 ctcagtccat tcacgatata gtgcatatcc ctttcgaatc ttgccatctg aatagccagc
      841 ttccagttga tgtcaatcag gtaggactcg attttgttca gatagttgaa ctcgaccaga
      901 tctcgaatct ttttgtactc agacacgcgg ttatagtcct tttcgaagga tttgtagttc
      961 ttattctgga tgttctttgc aaagaaatcg tcgttctcct tcagcttttt gatgtacttc
     1021 tccttgtatt cctttgagta gccgttcagt ttatcgttca gattcttcag aatagcgtcg
     1081 atctcactga ttttgttctt tctgatgttc tttccatcga tattaaacag gaatttagca
     1141 tcggccatct tgatgtcgtt gctaatcagg ccgtagatct tgtcaaagtt aggatttccg
     1201 atgttcagga acagattctt tttgtagata tacagctcgt tcttccgttc cttgggatag
     1261 tagatctcct ggaacttgtt ctcattttca gactccatat cctcaatcag attgtcgatt
     1321 tccttcttgt actttttcag aaagtcactg ttgaaaatga tcctgcacag gattttagac
     1381 ttgatctcct gatctttgtc cttgatatac tgatccacct ttttcttcag gtccttcttg
     1441 ttgatgttag acagctttct ctgttcatcc tgcaggatgt tggatttctt gtcgatctcg
     1501 aacttggttt catcgtcgaa gatgacaatc ttctccagtt tcttttccag cacatcacag
     1561 ccgttgatat cgtctttaaa ctcggtcaga atgttgttct tgatgtcctc gtagtagttg
     1621 ttgaagattt ctttcttagt ctggatcttg aagtcgtcga aatccttttc gatctctttc
     1681 atcttctgaa tgaactcttc caggttcaga ttccagttct ctgtgatgca ttcgttccgc
     1741 agggtattca gctgcataat ctcatccagg atgtcaatga tattctggta ttctgaggtg
     1801 ttcagccaca cgctggtggc gaagaatctg tttcggatct tattaatcac ggcattactg
     1861 ttcagcagtg caaagataga aatgatgtac tcgaaatcgt cgttgatgac aatggttttg
     1921 tcgctggttt tcacggtgat ccactcataa gtcttgttat cgttgatgtc cttaatctgt
     1981 ttcttgattt cctggatgtt catcttgaag tctgaaaagt cgaacagctc ttcgtagttt
     2041 ttgcgcagat atccaatgta gcactcgatc actttcttct gatatttctt gattgccttg
     2101 ttgttcccct tactggcaga aatctgtgcg ttcttgtagt agttctcgat gatgttctca
     2161 tcgatttcgt caatgttgcc cagggttttc ttcagttcct gcaggaagat attcttggac
     2221 tcgttctctt ccagatcgtc ctccaggatc agtttcttat acagctcctt attgacgtag
     2281 atcagagcgt tcagcacaat tttttcagtc tcgattgtgt caaaaggctc attcttggga
     2341 ttgttgcggt acaggttcag gatttcgggc agcactttgc tgaagctggg cagatacttg
     2401 atatcgttgt tgttctcttc agagatcttg atgtcattga tctttgtgat gatgtttttc
     2461 ttgtccttaa agaccacgtc caggttcaga gctttgctca cctcttcatc ggagatcttc
     2521 agattctgaa tgatgttgat cactttgttg taatcgtcct gagtcccctg caggtccctc
     2581 tctttggaaa tggcatgcag gatgcggttc ctttcattag tgccgatctt tgtaaacttt
     2641 cgaatgaagt tgtttgtgat gttgttcttg ttatcgatga aatccaggtc gcggatgatc
     2701 ttgatcttac tattcaggat tttcttatcc agcacatagt tcttctcccg gtcgcctcca
     2761 aagaaatcga tgttctcgtc gttgttgatg ttttctcggg aaaagatctt gttcagctcc
     2821 atatttgtgc tggcaaagaa ggtgatcagt tccaggtcca gctcttcctt ggcgtgcagg
     2881 cggctgaaat cgtcagtatt cactgtggtc atatcgatgt cgttatgcct cagcttcccc
     2941 aggtacataa tgtgctccag tgtatactgt ttcactctct tcagaatttt ctctgacagg
     3001 atactttcat tcagaatctt ttcgatctcg atcttctcca ttttcttcag tctgaccttc
     3061 tgctcgttca ccaggatttt ttcaattctg cccttcaggt atctgtagat gatcttatac
     3121 agctccttct cttcatcaga tttcttggaa aatttcttgc tgtcgaagtt caccttgtaa
     3181 tgcttcttaa agattccgaa gatctcggta tcacagttcc ctttcttcag ttccttctcc
     3241 agtttcttaa tcagctcgtc gatcttgaac tcagccagga tcttctcaat cttttccttg
     3301 atagagttgt tcttaatgtt ctcgacaaag aacttcacga tcttatcctt cttgttttct
     3361 ctctcgattt taaacttttc gtgcttgtcc agcagcacat aggacttgat gtatgtccga
     3421 tttctccgct tctccaggaa ctcgttattg actttcttca ctttctcaat gcgctttgtg
     3481 atgttccaaa attccagctc cttgatcacg aaatcggcaa tgtcctcgac ggtcagatcc
     3541 acattgatgt tcagaatctt ttccaccagc attttcttat ttttactttt ctttttgtcc
     3601 ccgccgacgt tcaggtaaaa cttcacgaag cccaggatct ccagattaga cttgatcttc
     3661 tcccggattt ccatgaagtt ggtcaggatc acgtcaatct tatcgtcttt cagcagcttc
     3721 tccctcaggt gctcttcgta gtatctattt tcaaacacct tttctgtctc gttttcgatg
     3781 atcttctcga tgatcttata caggctcata ttgatgttct tgaagatctc gtagatgctc
     3841 ttctttgtct ccagttcatc gttttcaatg atgcgcagaa tgatggagca gtcgttcagg
     3901 gtcttattag tgtactcatc ccggatgtca atttcgatct cttcctcgtt ctcctgtcgc
     3961 ttgatctcga tctttttatc gtccttagta atgccctgcc tgatggcctc gtcaatgatc
     4021 tttttctttg tgatccccag tgctttcagc ttctcgctct tgccgtaggc ctcgatatac
     4081 agcaccactt cctctgtctc caggaagtcg tcgttattct caattctaat gatcccttct
     4141 ttgcccttca gtttgaacag gatatttcct gcatgaaatt tcctagtgaa ctccttcaga
     4201 atattatcgt ttttcttgta gttgatgtac tttctgatga acttgttgtt gtcgatcttc
     4261 tctttgttgt tgttttcgtt gatgttcaga atgtacttgt tcccgtcata attccgcttg
     4321 actttcacct ttcgcttgat cttgaagtct ttcttatctc gcacctcgta ccaccgcttg
     4381 tgtccgaaca ggttccccat ggataagacc tcctaacata cgagccggaa gcataaagtg
     4441 taaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     4501 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     4561 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     4621 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     4681 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     4741 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     4801 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     4861 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     4921 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     4981 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     5041 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     5101 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     5161 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     5221 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     5281 cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag
     5341 ctgaacagga gggacagctg atagaaacag aagccactgg agcacctcaa aaacaccatc
     5401 atacactaaa tcagtaagtt ggcagcatca cccgacgcac tttgcgccga ataaatacct
     5461 gtgacggaag atcacttcgc agaataaata aatcctggtg tccctgttga taccgggaag
     5521 ccctgggcca acttttggcg aaaatgacac gttgatcggc acgtaagagg ttccaacttt
     5581 caccataatg aaataagatc actaccgggc gtattttttg agttatcgag attttcagga
     5641 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     5701 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     5761 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     5821 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     5881 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     5941 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     6001 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     6061 aaagggttta ttgagaatat gtttttcgtg tcagccaatc cctgggtgag tttcaccagt
     6121 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     6181 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     6241 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     6301 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac
     6361 gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg
     6421 tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt
     6481 tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc
     6541 aggctctccc cgtggaggta ataattgacg atatgatcat ttattctgcc tcccagagcc
     6601 tgataaaaac ggttagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     6661 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gcttgtttcg gcgtgggtat
     6721 ggtggcaggc cccgtggccg ggggactgtt gggcgctgcc ggcacctgtc ctacgagttg
     6781 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     6841 ggagctaccg gcagcggtgc ggactgttgt aactcagaat aagaaatgag gccgctcatg
     6901 gcgttgactc tcagtcatag tatcgtggta tcaccggttg gttccactct ctgttgcggg
     6961 caacttcagc agcacgtagg ggacttccgc gtttccagac tttacgaaac acggaaaccg
     7021 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     7081 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     7141 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     7201 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     7261 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     7321 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     7381 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     7441 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag
     7501 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     7561 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     7621 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     7681 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     7741 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     7801 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     7861 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     7921 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgacgctctc
     7981 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     8041 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     8101 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     8161 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     8221 cggccacgat gcgtccggcg tagaggatcc acaggacggg tgtggtcgcc atgatcgcgt
     8281 agtcgatagt ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg
     8341 acagtgctcc gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca
     8401 cgccatagtg actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta
     8461 ccggcataac caagcct
//
