LOCUS       sequence-181454-        7478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LbfCas13a 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 7478)
  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 7478)
  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: pC028 - LbfCas13a from Listeriaceae bacterium ...
FEATURES             Location/Qualifiers
     source          1..7478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(2819..2824,2825..2842,2843..2848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      2886..3431
                     /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     3332..3351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3548..3565
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3957..4059
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4060..4719
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4126..4145)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(5753..6943)
                     /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     6572..6591
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(7092..7126)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 ccctccttct tcagaaatgt atccagcttc ttgatagact tcaggcgatt gtcgccccag
       61 tactccaccc tctcggtgat gccggagtgc ttgaaatact ggttggccat gcactgaaaa
      121 tcgcggtcgg cgatggccac atagcccacg tacctgccca ggatctcgat cagcaggtgg
      181 tgcagctcgt tcacgtaggc cagtgtggtc ttattcttgt gccagttata ggcatcgatg
      241 gcgtcacagc actccttgta ctccttaaag aaggcattgt tctgaggctt cttgggattc
      301 ttggcccact cgttgtgcag ctccttgcgc ctcttgattg tctcctcgat tgtctccttc
      361 tggcgctccc actcggccag attacacttg ctcaccttga actcggggtt ggcatcaaac
      421 agcctctgga tcactgtctc ggtgccgtac ttccttgcct gctccactcc cctgaacagg
      481 attggtgtct tgccgtcaga ctgtctataa ggctcccgct ggtacagctc ctccaccaca
      541 tagcccttca tgatcttctc gtactctctc tcgctagaaa agaactgctt ccaatcgttc
      601 tcccgggagt ccactcccag cagggycagg ccgatgatct cctgctcgcc ggtcaggtgg
      661 ccatcgtact tgatcatctc atttctcagc tcgctcaggt gccttgcgtc cagcatcttg
      721 cagaagatcc aaaaggcgat ataatcgtcc ttgttcttca ggaaagaagg ctccaccttg
      781 atatcaaaag acatctcctc gatctcggac tgctccaggc cctgattgcg ggggttcttg
      841 atgaacttca gattggcaga gcgcatgtgc ctgtcgaagc ccttgataaa cacctgggag
      901 atgaacttct caaagtggtt tggggtatcc tcgatgctgt cctcgtggat ggcgatctgg
      961 ccctgcacat agcccatgta atctctaggc atctcgccct cctccacctc ccggatattg
     1021 ataaaggcct tggcgtcctg ctggcgattc ttgttcagcc tcagcacgaa gttggcggca
     1081 tccctaaagg cattgttgtc ggctgtgaac cagggcataa actgctgata gtacaccagc
     1141 ttggtcagga aatatctggc ccgaaaatcc tcctcgccgt actccaggat atcgaatgtg
     1201 gtcaggtcca gattatagta tctgttcttg gaggtctgat agtcgtggcc cttggcgtac
     1261 acccgcttga agctgggcac aaaaggggcg tttgtggtca gcagggaaaa gccctgcttc
     1321 atctcccaga tgctctgcag ctcgtcgtgt ctgtagaacc gcaccacgcc ggagctcttc
     1381 agcttctcct cgataaactc ggggatcttg gcgatatcct tgtccatcag cttctgcata
     1441 tcgctgtcgg cgtacttcat attgtttggc tcgaactcaa agcccttgat cttcaggatg
     1501 ctcttccaag agtgcttctt gcagtgatac acctcatttc tgatctgctg gatggagccc
     1561 cggatggccc agatggcctc atcgaaattc acgttctgca gctcgtcgct aaagaagggg
     1621 atcatcttct tcaccacatc agagcgtgtg gtggaatttg gcaggttctt cttgaaattg
     1681 cccttgccca ggatgtcctt ctcacactca gggttcagga tcatcttcag attgttggag
     1741 gcaaaggcgc aggcgttgat gaacttaaag ctgaaggcct cgccagacct gatatcctgc
     1801 aggtcctttg tcttgggatc ggtcagctcg taggcctcca tcttgccctg ctccagcaca
     1861 tagtggtaca cggcattctt gaactgctgc tccacggtgc tctcgatcag ctccttcttc
     1921 agaaagtcgg ctctggcgtg cttagagcgg ttgggggccc tcttgattgg gaaatacctc
     1981 tggatgtact tcttcacctc gatgccgtac tgattcagct cggggtcctc cttcagctcg
     2041 tccaggatgt tccgctggtg ctgctgcaga caggccttga tcttataatt ggcgctctta
     2101 tcctcgttca ggtccaccag cttgaaagac tcggccagat cggccagctt gtacttcttg
     2161 gccaccaggt ccaggcccag tggctctcca ttcagaaagg cggtcacata cttctgcagc
     2221 accagctttc tgggggagtc tgggctctgg atcttgttct gctcgataga cttctggatt
     2281 gtggtctgct tatagtggat gtaggactcg gcccaatctg tgtatggctt cagcttctca
     2341 aagaagatgc cccagtcagg cttgatctca tccaggtccc ggattgcggg cacggcgatg
     2401 tcggtcaggc agaactgcac gtgctctgct cccttgctag gtggataaga gatcatattc
     2461 tggaactttg gcttaaacag ctttgtcagg gactcggtat tcaggctctt caggttgtgc
     2521 agcttctggc taggcacctc ctggtgcagc acggccagca tcagctctgt gatctcctta
     2581 ttgatggcct tgttgatggc ctgctttctc cgattcttct ccttctcatc aggctttctc
     2641 acggtcttgt tcaggatgga cttgtagaag ctctccttct tcttatcaaa gatgatctct
     2701 gtggtcttgt tgccgtcgat gccctcatag cccagctggc cctccttgga tgtccgctcc
     2761 atcacgatgg ttctgccgtc cacccgcatc ttggtgatct tcatggataa gacctcctaa
     2821 catacgagcc ggaagcataa agtgtaaatc tagaaatatt ttatctgatt aataagatga
     2881 tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc
     2941 ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg
     3001 cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac
     3061 ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg
     3121 tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac
     3181 ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag
     3241 gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag
     3301 gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg
     3361 tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga
     3421 gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc
     3481 atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac
     3541 cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg
     3601 acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca
     3661 gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggcg gtgcttttgc
     3721 cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa acagaagcca
     3781 ctggagcacc tcaaaaacac catcatacac taaatcagta agttggcagc atcacccgac
     3841 gcactttgcg ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct
     3901 ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg acacgttgat
     3961 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     4021 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     4081 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     4141 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     4201 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     4261 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     4321 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     4381 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     4441 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtgtcagcc
     4501 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     4561 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     4621 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     4681 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     4741 tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga
     4801 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt
     4861 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
     4921 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga
     4981 tcatttattc tgcctcccag agcctgataa aaacggttag cgcttcgtta atacagatgt
     5041 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     5101 gggcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
     5161 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     5221 agtcatgccc cgcgcccacc ggaaggagct accggcagcg gtgcggactg ttgtaactca
     5281 gaataagaaa tgaggccgct catggcgttg actctcagtc atagtatcgt ggtatcaccg
     5341 gttggttcca ctctctgttg cgggcaactt cagcagcacg taggggactt ccgcgtttcc
     5401 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     5461 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     5521 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     5581 ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     5641 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     5701 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg
     5761 aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg
     5821 cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga
     5881 cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct
     5941 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     6001 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg
     6061 ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc
     6121 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     6181 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     6241 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     6301 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     6361 tcaagggcat cggtcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     6421 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     6481 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6541 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6601 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccacagga
     6661 cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga
     6721 ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca
     6781 tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga
     6841 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
     6901 tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc
     6961 gttagcaatt taactgtgat aaactaccgc attaaagctt gttttagtcc tcttttgttt
     7021 tgaggtctct aaatccgaga cgaatctccg tctccgtttt agtcctcttt tgttttgagg
     7081 tactctaaat cgctagcatt atacctagga ctgagctagc tgtcaatcac atctccagca
     7141 gtctcttcac gatgccacaa tactcctcgc tcacctggtt tgtctcgatg tagccgccat
     7201 cgatcttctt gccgcccagg tggcgcagct tcttaggctc caggctcttg atgaccagcc
     7261 ggtgcttatt ctccttcagg ttggcgaaca ccacggacat gccgtgtcta tccaggatgt
     7321 cgatcagaga cttggacacg gcattcttca gcttccggtc gtacttaaag atctcgcgca
     7381 gcctctcgga cagatacagc agggtacact cagacttcag ggacaggtag ttcaggtggg
     7441 cgatgtgatt ccgggcgttc ttctcgctca cgaacagg
//
