LOCUS       sequence-183041-        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LseCas13a 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 7686)
  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 7686)
  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: pC020 - LseCas13a from Listeria seeligeri
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(232..266)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        complement(join(3644..3649,3650..3667,3668..3673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      3711..4256
                     /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     4157..4176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4373..4390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4782..4884
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4885..5544
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4951..4970)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(join(6578..7686,1..82))
                     /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     7397..7416
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga
       61 tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata
      121 aactaccgca ttaaagcttg ttttagtcct ctttcatata gaggtagtct cttaccgaga
      181 cgaatctccg tctccgtttt agtcctcttt catatagagg tagtctctta cgctagcatg
      241 atacctagga ctgagctagc tgtcaatcac ttcatggtca gcagagttct caccagctga
      301 cagtattcgt tactgacctg gttgcttgac actgtagact tctcccccag atggtggatt
      361 ttcttgggct gcagtttgtc aatcttcaga tggtggttag tctggtacag tggcttgaag
      421 gtcacctcca tgccatgact agacaggatt tctttcagag acttggagac agcattcttc
      481 agtttcctgt catagctcag cacatctctg gcatcgtcga acagttccag aattgagttc
      541 cccagctggc cattcaggta gttaaagtga ctgatattgt tccgcttctc tttcagtcga
      601 ttgtcgaaca gttccagata ctccagggtc agccgcagct gcttcaggtc gactttcagc
      661 tggggatcat tcttggagct cactttgatg gaggcattct tcaggttata cagttcataa
      721 tcgttgtact tgttcttgtt cttgttctcg gacagcagaa tccagcttgt cactcggtat
      781 tcgctatttt cgcgctccag aatgtagttt gaactaaact ggaaatctct gtcagcgata
      841 gacatgtatc cggccagccg ggacagcagg tcaatggtca gctgatgcag gtgtctgacc
      901 tgggtcagct ccactttagt ctttgcccac tggtagttgg agatgtcatt aatcacgttc
      961 tggtatttct tagtccatgc gctatcccga gccagccctg gcttctcgat ccactgttta
     1021 tgcagggatt cctgctgagc aatcttttct gtcacgtcgt actcgtgcag cttggcaata
     1081 tcttttgcgc tcaccttata atcgtcagag gagctgaaca gcttttccag gatagtctct
     1141 gttccatatt tcttgaccag gtcgatactc cgattaatca caggtgtctg cccatcctcc
     1201 tgggtgtagg gcagagactg cagcagttcc tcgctcacat acagtgacat gtttttcttc
     1261 caggcttctt tatcgtcgaa cagctccttc cagtcattac atcctttctc cccgttcagc
     1321 agggccaggc cgatcacttc tcttgccttg gtaaaagttg aaatttcctc tgtactctgc
     1381 aggctacagc tgaacttgat catttcgttc cgcagctcgg acagctgctt tgcatccagc
     1441 agtttgcaca tcagccaaaa agcgatattg ctatcgtcca tgtctgtatg gaatggaatt
     1501 tcgatattat cgttctctgg cactgtgttc ttggtagggt ggcagatgta ggtcagccta
     1561 ttcttctcaa taaagctgtt gaaccctttg ataaacacct ggttaataaa cttttcgaag
     1621 tggttgattt tctccttttc ctcctgtttc ttctggtaca gcatcagctg actctgaata
     1681 taagacatgt attcggaagg tttctcgtcc ttattcattt tgcggatatc ctggaatgcc
     1741 ttagcatagc ccttcctctc tttgttcagg gtcaggataa agtccactga actcttgaac
     1801 agatcattgt ttgtggtaaa ctggggcagg aacacctgat agtacaccat cttaaacagg
     1861 gagtactggg cctgaaatgc ttcgctgttg aaggctttct cattgtagat gttcagcttc
     1921 agattatagt ctggctgtgc ctcatcctgg ttctggtagt cgaagccctt cagatagact
     1981 cttttaaagg aaggagcgaa gggcacggcg cttgtcagca gtgacagctc aaaattagga
     2041 atggtgaaca cctggttcag ctgatcagag gaatagtagt ccaggatctt gcttgactcc
     2101 attttgttaa tgatcagttc gggcacactg tccagctcag aataaaagta atccttgaac
     2161 agggtttcct tatacaggaa ctcgctggtc acgtctttgg tcttgccatt aatgatcttg
     2221 gatttcttca ctttgaaagt tgggttgtta aagaatttct tccagctatg tttcttcagg
     2281 tgaatgatct cgttccggat aggggcaatt gctcctcgca gtccccaaga ggccagctcg
     2341 atatcgtcca cagtaatttc ctgggaaaag aactggctcc actggcggat gaacttcttg
     2401 tgcttaatct ccttgaagct gttcttgaac tcgccgatca tcagaatgtc tttcttacag
     2461 acggggtaca ccatattcct cagattgtta gatgcgaaca ggcaagcgtt gataaacttc
     2521 agagcgaagg cttcctcaat tttgatcttc tgcagactat tagagttcac tgtagattcg
     2581 atctcatagg aggccagctt tccctcctgc aggattcgct ggacaatctt gttcttcagg
     2641 cggtggttca cgatcttctg aatttcgccg atctccagat tcctgatatc tcccactttt
     2701 ctgttggttt tcttaatagg gaagtaagtt tccagatgct tgcgaatctc gatattgaac
     2761 tggttcagtt cgctattctc tttcagttcc tcaatgatct gcctctcgtg gttctgcagg
     2821 gtagatttca gcttgtaatt gatctccttg tcgtttttct tagcttcttt aaaagtagcc
     2881 ttcagggcgg atgtcagctt ttccagctgg tatttctcgg acacgctctg gatatcgaag
     2941 ggcttgttct tgttcagaat atccttcata tagcggtcca tcagctggcc tgacctactc
     3001 tcggttgact ggatgcgatt gttcctgata cttttcttaa tcagtttagt cttactagag
     3061 atgtagttct cggcccagtt aatatacagt tccatgtcct tagaaaagga ctcccagcag
     3121 atgaaggttc cctgctgttt cttcagactg tcctccagca gctgagacag attaatttcg
     3181 attctgtatt cctctgactt atcgggactg ttctctggga aatagtacag gctcttcttg
     3241 aatctgtgat tcaggaagtt tgagatattc agcttagtga catcgctcac cttgatcttt
     3301 tcctggctgt tctcctgcag caggccgtgc accagtttct tcatctgctt ctgctcaggg
     3361 cgacaaatag tcttgttgac cacgcttttg tagaaggagc ttttcttatg gtgcatgatc
     3421 tgctctgtat tgtcctgcat ttcgttctca tagaccagtt tcccgccatt cttgtcccgg
     3481 ctgatcagca ctttcttccg atcgacctcc accttagtga tcctcattgt tttgacttcg
     3541 atgattttct tctctctccg gttatacatg taatcgcaaa agaacagcac gcccagatgg
     3601 tggatcaggg tcttaatgct gatccacatg gataagacct cctaacatac gagccggaag
     3661 cataaagtgt aaatctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt
     3721 tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt
     3781 tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag
     3841 tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc
     3901 tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga
     3961 ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat
     4021 acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca
     4081 aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca
     4141 cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4201 actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg
     4261 gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc
     4321 tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc
     4381 agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc
     4441 cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa
     4501 gccagtatac actccgctag cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc
     4561 cgtcagtagc tgaacaggag ggacagctga tagaaacaga agccactgga gcacctcaaa
     4621 aacaccatca tacactaaat cagtaagttg gcagcatcac ccgacgcact ttgcgccgaa
     4681 taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat
     4741 accgggaagc cctgggccaa cttttggcga aaatgacacg ttgatcggca cgtaagaggt
     4801 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     4861 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     4921 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     4981 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     5041 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     5101 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     5161 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     5221 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     5281 tatttcccta aagggtttat tgagaatatg tttttcgtgt cagccaatcc ctgggtgagt
     5341 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     5401 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     5461 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     5521 gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct
     5581 ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat
     5641 tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt
     5701 ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc
     5761 agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcatt tattctgcct
     5821 cccagagcct gataaaaacg gttagcgctt cgttaataca gatgtaggtg ttccacaggg
     5881 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg cttgtttcgg
     5941 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgctgccg gcacctgtcc
     6001 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     6061 ccaccggaag gagctaccgg cagcggtgcg gactgttgta actcagaata agaaatgagg
     6121 ccgctcatgg cgttgactct cagtcatagt atcgtggtat caccggttgg ttccactctc
     6181 tgttgcgggc aacttcagca gcacgtaggg gacttccgcg tttccagact ttacgaaaca
     6241 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     6301 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     6361 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     6421 ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     6481 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     6541 gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
     6601 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg
     6661 ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa
     6721 tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt
     6781 catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga
     6841 gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc
     6901 ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     6961 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     7021 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     7081 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     7141 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     7201 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     7261 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     7321 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     7381 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     7441 cggtgatgcc ggccacgatg cgtccggcgt agaggatcca caggacgggt gtggtcgcca
     7501 tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg cggcggccaa
     7561 agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg
     7621 ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc
     7681 ccggca
//
