LOCUS       sequence-193532-        7790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LwCas13a locus into pACYC184 with targeting spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7790)
  AUTHORS   Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung 
            J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, 
            Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, 
            Collins JJ, Zhang F
  TITLE     Nucleic acid detection with CRISPR-Cas13a/C2c2.
  JOURNAL   Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
  PUBMED    28408723
REFERENCE   2  (bases 1 to 7790)
  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: "Science. 
            2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321."
COMMENT     Sequence Label: pC011 LwCas13a locus with targeting spacer
FEATURES             Location/Qualifiers
     source          1..7790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     769..788
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(1298..1332)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(1336..4791)
                     /codon_start=1
                     /product="Leptotrichia wadei CRISPR-Cas effector that acts 
                     as an RNA-guided RNAse (Abudayyeh et al., 2017)"
                     /label=LwCas13a
                     /note="previously called C2c2"
                     /translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY
                     IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY
                     DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN
                     NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK
                     YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD
                     KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL
                     NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE
                     NENDITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE
                     IEDFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK
                     IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW
                     SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ
                     KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE
                     SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY
                     TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF
                     ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK
                     IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI
                     GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE
                     IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIRNY
                     IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK
                     IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     promoter        complement(join(4806..4811,4812..4829,4830..4835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      4873..5418
                     /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     5319..5338
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5535..5552
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5944..6046
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             6047..6706
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6113..6132)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(join(7740..7790,1..1140))
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
        1 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
       61 ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc
      121 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
      181 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
      241 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
      301 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
      361 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
      421 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
      481 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
      541 gactgggttg aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat
      601 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
      661 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
      721 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
      781 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      841 gtagaggatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag
      901 tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg
      961 tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat
     1021 gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     1081 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     1141 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttgtt
     1201 ttagtcccct tcgtttttgg ggtagtctaa atctatggat tacttggtag aacagcaatc
     1261 tgttttagtc cccttcgttt ttggggtagt ctaaatcgct agcattatac ctaggactga
     1321 gctagctgtc aatcattcca gggccttgta ctcgaacatc actttcacga gttcgcacag
     1381 ttcctcgctg ttccggtcgg tcatcagttt ctttttcttc agattcttca ggtgcacgat
     1441 cttctctgat tccagggtct ggatttcgat cttcttgtca gcgccgatct tgaaggtggc
     1501 cacgaagccg tattctttca gaatgtccac gatggacttc atgatggcgt tcttcagctt
     1561 ccggtcgtag gacagcagct tccgcaggtt ttccagcact tccagcaggc taatctcggc
     1621 gtgggggatg tagttgaagt gggcaatgta gttccggatg tacaggtcct ttttttcctg
     1681 cttcagtttc ttcactttct tgtcggagta gatgctccgc ttttccacat tgtccttgta
     1741 cagttctttg tagaagttga tatacttttc cacgatctgg ccgcttttgt acttcacatt
     1801 cttggagttg tcgaaattga aaatttcctc gatgtagtgg ttctcgggaa actcgccctt
     1861 cagccggaat ctcaggtccc gctcccagat gctggtgtag cccacgagcc ggtgcaggat
     1921 cttcagcagc aggccctgca gcaggttcag ctcattgaat tccaccttgt tcttcaggtg
     1981 ggtgtacttc tggatgttgc cgatggcctt ctcatactct ttgtagtcct cgtcgttgaa
     2041 cttttcgtcc ttcttgggtc tggcgtactt ccggtgcagg ttctgctgca tggtgtagtt
     2101 cttttcaatc tcattcttct tgttgctgta ctctttcagt tctttcaggc tgatcttata
     2161 cttggcctta tcggcgatct tttccagcag attcagcatg ccgtatttct tgatattgta
     2221 gaaggcccgg tgcttgatga tgttctcgcc gtcgaaatag atcttgttgg tgtcgaactt
     2281 tttcagctct ttccggtcct tgattttgtt ttcgttgaag tccaggaact tgccgatctc
     2341 gttggcttcc agctcgaagt cctcggtcac tctgttgttg tccaggttca gcaggttgat
     2401 cagttccagc tcgtcgctga aggtttcttc tttgttggcg gactggtact tttccaggct
     2461 gcccttcagg ttggtcagct ctttgtggtt cagcagcttc aggatcaggt aaaacatgtt
     2521 cagattctcg gtgtacttca gaatcttccc cagcttgatc tcgcgcacga actcattgat
     2581 ctcgtgaggg atttctttgt tccgattgtg cttctcatag ttcttcagga tcttgtcgta
     2641 cttctctttg ttatcctttt tgatcttgat cttggagaag atgtcgttgt tgtcattgtt
     2701 gttgttgctc tcgatatact tcagattgtt cttgttcagg tagtcgatga agcccttcag
     2761 gaaaatctgc tgaataaagt cgatgtaggt attcttttcc tctttgtcct ggttgttgat
     2821 catctctctg ctctggatga tggccaggta ttccacgggc acggttttct cgatgttctc
     2881 gaacttctga tacttgtagt ggccggtttt ctggttccgc tgcttgttaa tcttgatcac
     2941 ttcattggtg atcttaaaga acaccttgga gtttttcacg aacttgttca ggaactcgcc
     3001 gtagtagata ttcttcagca ggtagatctg ggcgtccttc tcttctttgt ccttgggcac
     3061 gctccaaaaa aacttcaggg tattccgcag gtcctcaatc ttgttgtaca gcttggtgaa
     3121 gctgggcacg aaggggatgt ttttgttcac gaagttgaac ttggtattct tcaggtactt
     3181 gatgatcaca tccttctcgt agtagttgaa cacgttggcg ctgttcagct gcttgaagat
     3241 tttcagcttc agcttctttt cgttgatttc gttctgaaac atcttcttgg agatctcgct
     3301 gggggcgata ttcttgaagg cgaagatgtc cttgccttcc agttccaggt tgaagtgcac
     3361 gatgccgtgt ctgatgctgc tgatggcctc gtcgatgttg gcgaagaagt cctcgatctc
     3421 gttcttgttg tccatgttga agtcgtagct gtagaacatc ttcagatttt ctttcacttc
     3481 gttctgcttg ttctcattgt agatcttgtc cacctcgccg gacacgtatt tctcttcgcc
     3541 cttgttgttc ttcacggtct tgccccgcat ccggccggtg atatcgttct cgttctcggt
     3601 ttccaggatg ttcctcaggc tgaagtaggc cacgctggac acgccgatga tgtttctcag
     3661 gaaggcctcg ttctgccggt tccgggcgat aaagtcggag gtggcgatct cgcccacttg
     3721 cagatagtag ttgtacttgc cgcagttccg cacgtaggtg tccagcttgt tcagcagttt
     3781 gttttcgatc agctttttca gattctggta ctcgaagatc cgcttgatct tatcgttgct
     3841 gatgttgctc agccgcttgt acacgtagtt tttcagcagc tgggacatct cgatttccac
     3901 gaagtggcag aaggcgtact taatattctt gtcgttcagt tcctctttgt ccaggtagta
     3961 cttgtagaac acctggcttt tcttcagctc agacatgtcg gggatcttct caatcagctc
     4021 tttgatgttg ttcacgttct ggatctcttc gtagataatc ttggcgaagt tctctttgtc
     4081 gttcttccgg ccgatgatct tgtgatagta ctcgcggatc ttgtacttct cgtgcttctt
     4141 gctgttctcg atcaggaaaa acagtttctc gatatcctct ttcttataca gcttgtcgaa
     4201 ggcttcctgc acgttgttga tgtagtcgtt gcgcttggcg ctctctctgt agtagtcgta
     4261 gatgatgttc cgcttgctct tgccgcccac tttttccacg ttgttctcgt tgatcttctg
     4321 gtagttggcc ttgttctctt cgaagctgta cttcaggctg ttgatcttgt tcagcttggc
     4381 ttccacgtcc ttccgaaaga tttccagttc ctcagagttc acgtcctcgt tcagcaggat
     4441 cttcttcagc acggagaagc tgttcttgtt tttcaggtcg tactcgctga tgtcctcttc
     4501 gctatagttc ttgtcctgca cggcgttctt ttctttccgg ttcttcagat acagcacgct
     4561 gtccttcagg tgcagcacct tgttgctaaa gaacttcttc aggttctctc ttctgatccg
     4621 gttctcttcc tcggaggcgt tgtcggggtt cttgatgtag atgtccagcc ggatgctcag
     4681 cagctcgctc agtctctcgc tggtccggtt ttcctcgctg gtggacttca cgagcttgcc
     4741 ctcttcgatg tacttcttgt ggctgatgcc gtcgaccttg gtcactttca tggataagac
     4801 ctcctaacat acgagccgga agcataaagt gtaaatctag aaatatttta tctgattaat
     4861 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     4921 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg
     4981 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc
     5041 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt
     5101 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5161 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga
     5221 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga
     5281 aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat
     5341 agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg
     5401 gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct
     5461 tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc
     5521 gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat
     5581 tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc
     5641 ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg
     5701 cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca
     5761 gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc
     5821 acccgacgca ctttgcgccg aataaatacc tgtgacggaa gatcacttcg cagaataaat
     5881 aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaca
     5941 cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     6001 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     6061 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     6121 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     6181 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     6241 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     6301 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     6361 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     6421 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     6481 gtcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     6541 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     6601 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     6661 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     6721 ttattggtgc ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa
     6781 tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg tcgttaaata
     6841 gccgcttatg tctattgctg gtttaccggt ttattgacta ccggaagcag tgtgaccgtg
     6901 tgcttctcaa atgcctgagg ccagtttgct caggctctcc ccgtggaggt aataattgac
     6961 gatatgatca tttattctgc ctcccagagc ctgataaaaa cggttagcgc ttcgttaata
     7021 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     7081 tggtgcaggg cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     7141 tgggcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     7201 cgacgatagt catgccccgc gcccaccgga aggagctacc ggcagcggtg cggactgttg
     7261 taactcagaa taagaaatga ggccgctcat ggcgttgact ctcagtcata gtatcgtggt
     7321 atcaccggtt ggttccactc tctgttgcgg gcaacttcag cagcacgtag gggacttccg
     7381 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     7441 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     7501 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     7561 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     7621 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     7681 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     7741 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
//
