LOCUS       sequence-181262-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LweCas13a 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 7236)
  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 7236)
  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: pC027 - LweCas13a from Listeria weihenstephane...
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1507..1512,1513..1530,1531..1536))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      1574..2119
                     /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     2020..2039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2236..2253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2645..2747
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2748..3407
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2814..2833)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(4441..5631)
                     /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     5260..5279
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(5781..5815)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 agttggcggc atcgcgaaag gcattgttgt cggctgtgaa ccaaggcata aactgctgat
       61 agtacaccag cttggtcagg aaatagcgtg cccggaagtc ctcctctccg tactccagga
      121 tatcgaatgt ggtcaggccc aggtcatagt agcgattctt gcttgtctga tagtcgtggc
      181 ccttggcgta caccctcttg aaggatggca caaaaggggc gtttgtggtc agcaggctaa
      241 agccctgctt catctcccag atagactgca gcttatcgtg ggagtagaac ctgatgatgc
      301 cggagctctt cagcttctcc tcgataaagt cggggatctt ggcgatatcc ttgtccatca
      361 gcttctgcat atcggagtcg gtgtacttca tattgtttgg ctcgaactca aagcccttga
      421 tcttcaggat gctcttccaa gagtgcttct tgcagtgata cacctcgttt ctgatctgct
      481 ggatagagcc ccggatggcc cagatggcct catcgaaatt cacgttctgg atctcgtcgg
      541 aaaagaaggg gatcatcttc ttcaccacgt cgctctgtgt ggtagaattt ggcaggttct
      601 tcttgaaatc gcccttgccc aggatgtcct tctcacactc aggattcagg atcatcttca
      661 gattgttgct ggcaaaggcg caggcgttga tgaacttaaa gctgaaggcc tcgccagatc
      721 tgatatcctg caggtccttt gtcttgggat cggtcagctc gtaggcctcc atcttgccct
      781 gctccagcac atagtggtac acggcattct tgaactgctg ctccacggtg gactcgatca
      841 gctccttctt cagaaagtcg gctctggcgt gcttgctgcg gtttggtgcc ctcttgattg
      901 ggaaataccg ctggatgtac ttcttcacct cgatgccgta ctgattcagc tcggggtcct
      961 ccttcagctc gtccaggatg ttccgctggt gctgctgcag acaggccttg atcttataat
     1021 tggcagactt atcctcgttc aggtccacca ccttgaagga ctcggccaga tcggccagct
     1081 tgtacttctt ggccaccagg tccaggccca gtggctctcc attcagaaag gcggtcacat
     1141 acttctgcag caccagcttc cttggggagt ctgggctctg gatcttgttc tgctcgatgc
     1201 tcttctggat tgtggtctgc ttatagtgga tgtaagactc ggcccaatct gtgtatggct
     1261 tcagcttctc aaagaagatg ccccagtcag gcttgatctc atccaggtcc ctgattgcgg
     1321 gcacggcgat gtcggtcagg cagaactgca cgtgctctgc tcccttggaa ggtggatagc
     1381 tgatcatgtt ctggaacttt ggcttaaaca gctttgtcag ggactcggta ttcaggctct
     1441 tcaggttgtg cagcttctgg gaaggcacct cctggtgcag cagggccagc atggataaga
     1501 cctcctaaca tacgagccgg aagcataaag tgtaaatcta gaaatatttt atctgattaa
     1561 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     1621 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     1681 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     1741 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     1801 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1861 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     1921 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     1981 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     2101 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     2161 ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt
     2221 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata
     2281 ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac
     2341 cctcatcagt gccaacatag taagccagta tacactccgc tagcgctgat gtccggcggt
     2401 gcttttgccg ttacgcacca ccccgtcagt agctgaacag gagggacagc tgatagaaac
     2461 agaagccact ggagcacctc aaaaacacca tcatacacta aatcagtaag ttggcagcat
     2521 cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa
     2581 taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgac
     2641 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     2701 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     2761 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     2821 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     2881 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     2941 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     3001 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     3061 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     3121 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     3181 tgtcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     3241 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     3301 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     3361 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca
     3421 gttattggtg cccttaaacg cctggttgct acgcctgaat aagtgataat aagcggatga
     3481 atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat
     3541 agccgcttat gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt
     3601 gtgcttctca aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga
     3661 cgatatgatc atttattctg cctcccagag cctgataaaa acggttagcg cttcgttaat
     3721 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     3781 atggtgcagg gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     3841 ttgggcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     3901 gcgacgatag tcatgccccg cgcccaccgg aaggagctac cggcagcggt gcggactgtt
     3961 gtaactcaga ataagaaatg aggccgctca tggcgttgac tctcagtcat agtatcgtgg
     4021 tatcaccggt tggttccact ctctgttgcg ggcaacttca gcagcacgta ggggacttcc
     4081 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4141 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4201 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     4261 gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
     4321 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
     4381 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
     4441 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
     4501 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
     4561 cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc
     4621 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
     4681 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
     4741 cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg
     4801 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     4861 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     4921 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     4981 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     5041 gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca
     5101 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     5161 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     5221 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     5281 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     5341 ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc
     5401 gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag
     5461 aaattgcatc aacgcatata gcgctagcag cacgccatag tgactggcga tgctgtcgga
     5521 atggacgata tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc
     5581 atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc
     5641 ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttgt tttagacctc
     5701 ttctattttg aggtactcta aatccgagac gaatctccgt ctccgtttta gacctcttct
     5761 attttgaggt actctaaatc gctagcatta tacctaggac tgagctagct gtcaatcaga
     5821 tctccagcag tctcttcacg atgccacaat actcctcgct cacctgattt gtctcgatgt
     5881 agccgttatc gatcttcttc tcgcccaggt gtctcagctt ctttggctcc aggctcttga
     5941 tgaccagccg gtgcttattc tccttcaggt tggcgaacac cacagacatg ccgtggcgat
     6001 ccaggatgtc gatcagagac ttggacacgg cattcttcag cttcctgtcg tacttaaaga
     6061 tctctctcag ccgctcggac agatacagca gggtacactc agacttcagg gacaggtagt
     6121 tcaggtgggc gatgtgattc ctggcgttct tctcggacac gaacaggccc tccttcttca
     6181 gaaatgtatc cagcttcttg atgctcttca ggcgattgtc gccccagtac tccaccctct
     6241 cggtgatgcc agagtgcttg aaatactggt tggccatgca ctgaaaatcg cggtcggcga
     6301 tggccacata gcccacgtac ctgcccagga tctcgatcag caggtggtgc agctcattca
     6361 cgtacaccag tgtggtctta ttcttgtgcc agttataggc atcgatggcg tcacagcact
     6421 ccttgtactc cttaaagaag gcattgttct gaggcttctt gggattcttc tcccactcgt
     6481 tgtgcagctc ctttctccgc tcgattgtct cctcgattgt ctccttctgt ctctcccact
     6541 ctgtgatgtt acacttgctc accttgaact caggagaggc atcaaacagc cgctggatca
     6601 ctgtctcggt gccgtacttc cttgcctgct ccactcccct gaacaggatt ggggtcttgc
     6661 cgtcgctctg tctataaggc tcccgctggt acagctcctc gcccacatag cccttcatga
     6721 tcttctcgta ctcgcgctcg ctagaaaaga actgcttcca atcgttctcc ctagagtcca
     6781 ctcccagcag ggccaggccg atgatctcct gctcgcctgt caggtggcca tcgtacttga
     6841 tcatctcatt tctcagctcg gacaggtgcc ttgcgtccag catcttgcag aaggtccaaa
     6901 aggcgatata atcgtccttg ttcttcagga agctaggctc caccttgatg tcaaaggaca
     6961 tctcctcgat ctcgctctgc tccaggccct gatttctggg gttcttgatg aacttcagat
     7021 cggcagaccg catgtgggag tcgaagccct tgataaacac ctgagagatg aacttctcaa
     7081 agtggtttgg tgtatcctcg gtggagtcct cgtggatggc gatctggccc tgcacatagc
     7141 ccatgtaatc tctaggcatc tcgccctcct ccacctcccg gatattgata aaggccttgg
     7201 cgtcctgctg gcgattcttg ttcagcctca gcacga
//
