LOCUS Exported 7788 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses PpCas13a for bacterial expression and contains backbone for spacer cloning. ACCESSION . VERSION . KEYWORDS pC026 - PpCas13a from Paludibacter propionicigenes WB4 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7788 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(2583..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 //