LOCUS       sequence-183043-        7788 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses PpCas13a 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 7788)
  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 7788)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pC026 - PpCas13a from Paludibacter propionicig...
FEATURES             Location/Qualifiers
     source          1..7788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(2583..2588,2589..2606,2607..2612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      2650..3195
                     /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     3096..3115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3312..3329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3721..3823
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3824..4483
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3890..3909)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(5517..6707)
                     /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     6336..6355
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(6857..6891)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 gctcttctgg gctgtattcc aggcggtgaa gttggcctct gtggtcttaa actgggtgtc
       61 cttgttgatg atctgctcca gcagctttgt ggtgccgtac ttcacagaca gctcgatatt
      121 ggcgtggatc acaggagagt gcttatcgga ctgcacgaac aggtcggacc agtttgtgat
      181 atcggcgccc tgctcgataa aagggcgcag ccttgccaga caatctgcct cggagctata
      241 cagatctctg tagtcggtgg gcaccacgtc ggcgctcagc aggcagatct cgatgatctc
      301 cagcaggtgg tggaacttaa actcgttcac ggactctctg aacttgatca gctcgttccg
      361 cagattgctc aggtgggcgg cgtccagcag cttgcagaac acgtagaagg cgatgtgggt
      421 ggcatcgtcg gccttggcgt actggggtgt cagcttgcaa tcggcggtga tgctggcctc
      481 cagctgattc agcttgtcgg ccttctgctg gttagaggct gttgcggtca gctgaggctg
      541 tggcatctgc acgaaatcaa actccttggc gcgcaggaag gagtcaaagc ccttgatgaa
      601 cacctgcagc acgaacttct caaaattgat ccttgtctcc tcggccacct tctcctcctt
      661 cttgttctgc tcctgcatca gctcagactg cacataggcc atgtaatcgg cgatggagtc
      721 tgctgcggtc atggggcgca cggcctcaaa ggcgtaggcc tccttcttcc ttggattcac
      781 tttctcggcc tgcttcttgt tctgcatctg cacgaagccc acgctgtcgg caaaggcctt
      841 ccgatctgtg gtaaagcctg gcaggaacag attgttatag atcagcttca gcatgaaata
      901 gcgggcattg taggactcct cggcaaagtt ctccttcctc agatactgct ccagcatcag
      961 ctcgtagaag ctctcgtcct gcttggcgtt ctggtaattg atgccgccgt taaacacctt
     1021 cttgaagccg ggggcaaatg ggatggtgct gcggcacaga gagaactgaa atgtggtcag
     1081 caggctcttc agattctcga ttgtatagta agacacggct ccgccggttt tcagctgctg
     1141 ggcgaaggcc tcagggatct tctccagctc attgataaac ctggccttat agattgtatc
     1201 ggcgtagttg gtctgctgct tagggtctgt gatggtgggg ttctcgaaat tggagatgtt
     1261 gaacacggtt ttcagggcat ccttcttata gtggttcaca ttgttgcgga tctgctgcac
     1321 ggctcccctg atgccccaca gctgtgtctc cttctcggcc ttattggtgc tcaggccctc
     1381 gccgaaaaag aaggagtaca gctggctatt tgtgttgtcc ttcagcagag acttgatgaa
     1441 gtcgcccttg cccaggatat cattggtctg ctcgttgtcc accatatttc tgatattgtt
     1501 ggcggcgaag gcacatgtgc cggtcagatt cagcacaaag gcctcgtttg tcttgatccg
     1561 gatcagatca ttgcttgtgg tgtcggcctt cagctcgtgg tggttggtct tgccctgctg
     1621 gatgatattg gctctgatgg cgttcaccag ctgcttctcg attgtggttt tcagggtctg
     1681 ggccttcaga tagtgggcga tatcgtcggc tgtgtttctc cgcttagagg tcttgatggg
     1741 gaaatagtgc tccaggtact tcaccacctc caggtggtag atggacagct ggttatcggc
     1801 tctattggcc tcgtttggca cctcccgggt gccaaagatg gcaggctggt gattctgcag
     1861 ggccttcttc aggccccggt ggtaggcatt tgtgttgatc ttgcccttat cgtcggtcag
     1921 tggcttcaca tcctgcagca tcttgcacag cacgtctgtc atatacacct tgtggtagtg
     1981 ggtcaggtcg atggagccgc tggctgtgaa ctctgtctcc caggccagca gggccttctt
     2041 gcgcttagac aggttctctg tcagatcgat cctattgttc tcgatggact tcttcagcag
     2101 gtcgctcttt gtctggatat accaggtttt ccaatcatag tagggctgca gcttgcgctt
     2161 gtcctgtgcc tgcacggcct ccacgatcag cagggtcaga ttgaaggact cctccttgcc
     2221 gttcttccag tacttcacgg gctcctggaa cttgtgattc agaaatggca ggatctcctc
     2281 ggccttgatc tgctttggca gctcctgctt aggatactta aagatcttct ccacgatctt
     2341 gtactgatcc acattcagct tctgcttctt ggctgtgtcg aacttgatga ttgtcttgtt
     2401 cagggtagac aggtcaaagg actgcctctt cttctcaggc aggatgttgc ttgtctcatt
     2461 agacacgggc tgtccagagt acaccagctg tgcgcctgtg gtctttctgt gcaccagcac
     2521 catcttatcc ttgccgccgt ccttcacctt caccttggac acccgcatgg ataagacctc
     2581 ctaacatacg agccggaagc ataaagtgta aatctagaaa tattttatct gattaataag
     2641 atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
     2701 cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa
     2761 ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca
     2821 tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg
     2881 catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact
     2941 gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg
     3001 tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag
     3061 gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt
     3121 cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg
     3181 cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc
     3241 tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa
     3301 cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct
     3361 gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc
     3421 atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt
     3481 ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa
     3541 gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc
     3601 cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa
     3661 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgacacgt
     3721 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     3781 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     3841 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     3901 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     3961 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     4021 gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga
     4081 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     4141 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     4201 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtgtc
     4261 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     4321 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     4381 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     4441 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     4501 ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg
     4561 cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc
     4621 gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt gaccgtgtgc
     4681 ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat aattgacgat
     4741 atgatcattt attctgcctc ccagagcctg ataaaaacgg ttagcgcttc gttaatacag
     4801 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     4861 tgcagggcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     4921 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     4981 cgatagtcat gccccgcgcc caccggaagg agctaccggc agcggtgcgg actgttgtaa
     5041 ctcagaataa gaaatgaggc cgctcatggc gttgactctc agtcatagta tcgtggtatc
     5101 accggttggt tccactctct gttgcgggca acttcagcag cacgtagggg acttccgcgt
     5161 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5221 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     5281 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     5341 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     5401 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     5461 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     5521 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     5581 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     5641 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     5701 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     5761 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     5821 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     5881 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     5941 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     6001 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     6061 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     6121 gctctcaagg gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     6181 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     6241 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     6301 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     6361 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatccac
     6421 aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc
     6481 aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat
     6541 tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg
     6601 acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
     6661 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
     6721 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttgttgta gttcccttca
     6781 attttgggat aatccacaag cgagacgaat ctccgtctcc gttgtagttc ccttcaattt
     6841 tgggataatc cacaaggcta gcattatacc taggactgag ctagctgtca atcacttctt
     6901 catctccagc aggctccggc acatattgca ataggccatc atgacctggt tggtacaata
     6961 ctggtactct ggcttatcct tggcagagga gcccaggtgg tagatcttct taggctccag
     7021 agactccacc ttcagcttgt ggtcggcgtt cagcttcagc ttcaggatca tgccgtgctt
     7081 atcgaacagg tcgataaagg ccttggacac ggcattcttc agcttgcgat cgtagtgcag
     7141 cagctccctc agctcgttga tcaggtcgat caggctgaaa tcggcggcgt ccttggtcag
     7201 ataattaaag tgggcgatgt ggtttctgat atcgtacatc tgcttctcct tgatctcgtc
     7261 attgctcacg tgcaggaagg cattgttata gtagtttctc ttgctagaga taaactggat
     7321 cagattggcc cgcacagact cgttgatctt attgccgttg atctggatga tcttgttccg
     7381 gatatcgtag atctccttga tcttcttggt gggcacctca ttcagcttct tgtcgatgga
     7441 gtgcaggttc acgaagccgt gcagcttaaa ctcatcggcg atctgctgct cgtcgaaaaa
     7501 ctggaaatct ctgtcaaaca gggccacgaa tcctgccatc cttcccagca gctcgattgt
     7561 caggccgtgc aggcgattca ggtgcacgaa gtgcatcttg ttgtccagcc agttataggt
     7621 attgatgtaa tcgcagatgt ccagataatc gtcgccgtgc ttctcgataa acttctgggc
     7681 gaaattggac ttctctctct tccgctcctg cttctcctta tcgtcggcgt tcttggcctt
     7741 cacccactgc tcgtggtgat cctccctctg cttgatcagc tgctcgat
//
