LOCUS       sequence-181266-        8344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LbmCas13a 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 8344)
  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 8344)
  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: pC021 - LbmCas13a from Lachnospiraceae bacteri...
FEATURES             Location/Qualifiers
     source          1..8344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3441..3475
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             3623..4813
                     /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     complement(3975..3994)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5847..6506)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6421..6440
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6507..6609)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7001..7018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7135..7680)
                     /direction=LEFT
                     /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     complement(7215..7234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7718..7723,7724..7741,7742..7747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 agaatcagaa catggtcatc cagcccgata gcgactccca ggtgctgtct ctgagcgagt
       61 cccggagaga gaagcagtct agcgccgtgt cctctgacac cctggtgaat tgcaaggaga
      121 aggacgtgct gaaggccttc ctgacagatt acgccgtgct ggatgaggac gagcgcaaca
      181 gcctgctgtg gaagctgcgg aatctggtga acctgtactt ttatggctct gagagcatca
      241 gggattactc ttataccaag gagaagagcg tgtggaagga gcacgacgag cagaaggcca
      301 acaagaccct gttcatcgat gagatctgcc acatcacaaa gatcggcaag aacggcaagg
      361 agcagaaggt gctggactat gaggagaata ggagccgctg tcggaagcag aatatcaact
      421 actatcgctc cgccctgaat tacgccaaga acaataccag cggcatcttc gagaatgagg
      481 attccaacca cttttggatt cacctgatcg agaacgaggt ggagcggctg tataatggca
      541 tcgagaacgg cgaggagttc aagtttgaga caggctacat ctctgagaaa gtgtggaagg
      601 ccgtgatcaa ccacctgagc atcaagtaca tcgccctggg caaggccgtg tataactatg
      661 ccatgaagga gctgagctcc cccggcgaca tcgagcctgg caagatcgat gactcctata
      721 tcaacggcat cacctctttc gactacgaga tcatcaaggc cgaggagtcc ctgcagagag
      781 atatctctat gaatgtggtg tttgccacaa actacctggc ctgcgccacc gtggatacag
      841 acaaggattt cctgctgttt agcaaggagg acatcaggtc ctgcaccaag aaggatggca
      901 acctgtgcaa gaacatcatg cagttctggg gcggctattc cacatggaag aatttttgtg
      961 aggagtacct gaaggatgac aaggacgccc tggagctgct gtactccctg aagtctatgc
     1021 tgtattctat gagaaactct agcttccact tttctaccga gaatgtggac aacggcagct
     1081 gggatacaga gctgatcggc aagctgttcg aggaggattg taaccgcgcc gcccggatcg
     1141 agaaggagaa gttctacaac aataacctgc acatgtttta ttcctctagc ctgctggaga
     1201 aggtgctgga gagactgtac tcctctcacc acgagagggc cagccaggtg ccttccttca
     1261 acagagtgtt tgtgaggaag aatttcccaa gctccctgtc cgagcagaga atcaccccca
     1321 agtttacaga ctccaaggat gagcagatct ggcagtctgc cgtgtactat ctgtgcaagg
     1381 agatctacta taacgacttc ctgcagagca aggaggccta caagctgttt agggagggcg
     1441 tgaagaatct ggacaagaac gatatcaata accagaaggc cgccgatagc ttcaagcagg
     1501 ccgtggtgta ctatggcaag gccatcggca acgccaccct gtcccaggtg tgccaggcca
     1561 tcatgacaga gtataatagg cagaataacg acggcctgaa gaagaagagc gcctacgccg
     1621 agaagcagaa cagcaacaag tacaagcact atcctctgtt cctgaagcag gtgctgcaga
     1681 gcgccttttg ggagtacctg gatgagaaca aggagatcta tggcttcatc tctgcccaga
     1741 tccacaagag caacgtggag atcaaggccg aggactttat cgccaattac tctagccagc
     1801 agtataagaa gctggtggat aaggtgaaga aaacccctga gctgcagaag tggtatacac
     1861 tgggccgcct gatcaatcca cggcaggcca accagttcct gggctccatc agaaattacg
     1921 tgcagtttgt gaaggacatc cagaggcgcg ccaaggagaa tggcaaccca atcaggaact
     1981 actatgaggt gctggagtcc gattctatca tcaagatcct ggagatgtgc accaagctga
     2041 atggcaccac aagcaacgac atccacgatt acttccgcga cgaggatgag tacgccgagt
     2101 atatctccca gttcgtgaac tttggcgacg tgcactctgg cgccgccctg aatgcctttt
     2161 gcaacagcga gtccgagggc aagaagaacg gcatctacta tgacggcatc aatcccatcg
     2221 tgaatcggaa ctgggtgctg tgcaagctgt atggctcccc tgatctgatc tctaagatca
     2281 tcagccgcgt gaatgagaac atgatccacg acttccacaa gcaggaggat ctgatccggg
     2341 agtaccagat caagggcatc tgttctaaca agaaggagca gcaggacctg cgcacctttc
     2401 aggtgctgaa gaatcgcgtg gagctgcggg atatcgtgga gtacagcgag atcatcaacg
     2461 agctgtatgg ccagctgatc aagtggtgct acctgagaga gagggacctg atgtacttcc
     2521 agctgggctt tcactacctg tgcctgaata acgcctcctc taaggaggcc gattatatca
     2581 agatcaatgt ggatgaccgg aacatcagcg gcgccatcct gtaccagatc gccgccatgt
     2641 atatcaatgg cctgcccgtg tactataaga aggatgacat gtacgtggcc ctgaagtccg
     2701 gcaagaaggc ctctgacgag ctgaatagca acgagcagac ctccaagaag atcaactact
     2761 tcctgaagta tggcaataac atcctgggcg acaagaagga tcagctgtac ctggccggcc
     2821 tggagctgtt cgagaatgtg gccgagcacg agaacatcat catctttaga aatgagatcg
     2881 accacttcca ctacttttat gaccgcgatc ggtccatgct ggatctgtat tctgaggtgt
     2941 ttgaccgctt ctttacctac gatatgaagc tgcggaagaa tgtggtgaac atgctgtata
     3001 acatcctgct ggaccacaat atcgtgagct ccttcgtgtt tgagacaggc gagaagaagg
     3061 tcggcagggg cgatagcgaa gtgatcaagc cttccgccaa gatcagactg agggccaata
     3121 acggcgtgtc tagcgacgtg ttcacctaca aagtgggcag caaggatgag ctgaagatcg
     3181 ccacactgcc agccaagaac gaggagtttc tgctgaatgt ggccagactg atctactatc
     3241 ccgacatgga ggccgtgtcc gagaacatgg tgagggaggg cgtggtgaag gtggagaagt
     3301 ctaatgataa gaagggcaag atcagcagag gctccaatac caggtcctct aaccagtcta
     3361 agtacaacaa caagagcaag aacagaatga actactctat gggcagcatc ttcgagaaga
     3421 tggacctgaa gtttgattga ttgacagcta gctcagtcct aggtataatg ctagcgtatt
     3481 gagaaaagcc agatatagtt ggcaatagac ggagacggag attcgtctcg gtattgagaa
     3541 aagccagata tagttggcaa tagacaagct ttaatgcggt agtttatcac agttaaattg
     3601 ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac
     3661 cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg
     3721 ggatatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc
     3781 gttgatgcaa tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg
     3841 cccagtcctg ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac
     3901 acccgtcctg tggatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     3961 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     4021 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     4081 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     4141 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaccgatgcc
     4201 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     4261 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg
     4321 ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc
     4381 ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg
     4441 tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt
     4501 gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg
     4561 cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca
     4621 gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct
     4681 gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt
     4741 aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc
     4801 cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt
     4861 ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata
     4921 tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc
     4981 tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     5041 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
     5101 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
     5161 agtctggaaa cgcggaagtc ccctacgtgc tgctgaagtt gcccgcaaca gagagtggaa
     5221 ccaaccggtg ataccacgat actatgactg agagtcaacg ccatgagcgg cctcatttct
     5281 tattctgagt tacaacagtc cgcaccgctg ccggtagctc cttccggtgg gcgcggggca
     5341 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     5401 cggcagcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     5461 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
     5521 cacctacatc tgtattaacg aagcgctaac cgtttttatc aggctctggg aggcagaata
     5581 aatgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact ggcctcaggc
     5641 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     5701 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc
     5761 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa
     5821 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
     5881 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     5941 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     6001 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     6061 ggattggctg acacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     6121 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     6181 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     6241 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga
     6301 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     6361 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     6421 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     6481 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     6541 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     6601 tgccgatcaa cgtgtcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg
     6661 acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa
     6721 agtgcgtcgg gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc
     6781 tccagtggct tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg
     6841 taacggcaaa agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt
     6901 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
     6961 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
     7021 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
     7081 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     7141 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     7201 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     7261 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     7321 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
     7381 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7441 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     7501 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     7561 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     7621 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     7681 gaagatcatc ttattaatca gataaaatat ttctagattt acactttatg cttccggctc
     7741 gtatgttagg aggtcttatc catgcagatc tccaaggtga accacaagca cgtggcagtg
     7801 ggacagaagg acagggagag aatcaccggc ttcatctata atgacccagt gggcgatgag
     7861 aagagcctgg aggacgtggt ggcaaagagg gccaacgata ccaaggtgct gttcaacgtg
     7921 ttcaacacaa aggacctgta cgattctcag gagagcgaca agtccgagaa ggataaggag
     7981 atcatctcca agggcgccaa gttcgtggcc aagtctttta acagcgccat cacaatcctg
     8041 aagaagcaga ataagatcta ttccaccctg acatctcagc aggtcatcaa ggagctgaag
     8101 gacaagttcg gcggcgcccg catctacgac gatgacatcg aggaggccct gaccgagaca
     8161 ctgaagaagt cttttagaaa ggagaatgtg aggaacagca tcaaggtgct gatcgagaat
     8221 gcagcaggca tccggagctc cctgtccaag gacgaggagg agctgatcca ggagtatttc
     8281 gtgaagcagc tggtggagga gtacaccaag acaaagctgc agaagaacgt ggtgaagagc
     8341 atca
//
