LOCUS Exported 8410 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses FnCpf1 and spacers 1-4 of CRISPR array under artificial promoter. Cas genes are removed.. ACCESSION . VERSION . KEYWORDS pY002 (pFnCpf1_min) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8410) AUTHORS Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F TITLE Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. JOURNAL Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. PUBMED 26422227 REFERENCE 2 (bases 1 to 8410) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8410 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 134..153 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(220..322) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind complement(714..731) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(848..1393) /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(928..947) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 1426..1455 /label=lac promoter /note="promoter for the E. coli lac operon" promoter 5373..5407 /label=J23119 promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119)" CDS 5746..6936 /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(6098..6117) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(7970..219) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" ORIGIN 1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt 61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt 121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga 181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga 241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt 301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc 361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat 421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt 481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg 541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact 601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa 661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc 721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc 781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa 841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc 901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc 961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc 1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac 1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt 1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt 1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg 1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt 1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc 1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tattttttac actttatgct 1441 tccggctcgt atgttaggag gtctttatca tgtcaattta tcaagaattt gttaataaat 1501 atagtttaag taaaactcta agatttgagt taatcccaca gggtaaaaca cttgaaaaca 1561 taaaagcaag aggtttgatt ttagatgatg agaaaagagc taaagactac aaaaaggcta 1621 aacaaataat tgataaatat catcagtttt ttatagagga gatattaagt tcggtttgta 1681 ttagcgaaga tttattacaa aactattctg atgtttattt taaacttaaa aagagtgatg 1741 atgataatct acaaaaagat tttaaaagtg caaaagatac gataaagaaa caaatatctg 1801 aatatataaa ggactcagag aaatttaaga atttgtttaa tcaaaacctt atcgatgcta 1861 aaaaagggca agagtcagat ttaattctat ggctaaagca atctaaggat aatggtatag 1921 aactatttaa agccaatagt gatatcacag atatagatga ggcgttagaa ataatcaaat 1981 cttttaaagg ttggacaact tattttaagg gttttcatga aaatagaaaa aatgtttata 2041 gtagcaatga tattcctaca tctattattt ataggatagt agatgataat ttgcctaaat 2101 ttctagaaaa taaagctaag tatgagagtt taaaagacaa agctccagaa gctataaact 2161 atgaacaaat taaaaaagat ttggcagaag agctaacctt tgatattgac tacaaaacat 2221 ctgaagttaa tcaaagagtt ttttcacttg atgaagtttt tgagatagca aactttaata 2281 attatctaaa tcaaagtggt attactaaat ttaatactat tattggtggt aaatttgtaa 2341 atggtgaaaa tacaaagaga aaaggtataa atgaatatat aaatctatac tcacagcaaa 2401 taaatgataa aacactcaaa aaatataaaa tgagtgtttt atttaagcaa attttaagtg 2461 atacagaatc taaatctttt gtaattgata agttagaaga tgatagtgat gtagttacaa 2521 cgatgcaaag tttttatgag caaatagcag cttttaaaac agtagaagaa aaatctatta 2581 aagaaacact atctttatta tttgatgatt taaaagctca aaaacttgat ttgagtaaaa 2641 tttattttaa aaatgataaa tctcttactg atctatcaca acaagttttt gatgattata 2701 gtgttattgg tacagcggta ctagaatata taactcaaca aatagcacct aaaaatcttg 2761 ataaccctag taagaaagag caagaattaa tagccaaaaa aactgaaaaa gcaaaatact 2821 tatctctaga aactataaag cttgccttag aagaatttaa taagcataga gatatagata 2881 aacagtgtag gtttgaagaa atacttgcaa actttgcggc tattccgatg atatttgatg 2941 aaatagctca aaacaaagac aatttggcac agatatctat caaatatcaa aatcaaggta 3001 aaaaagacct acttcaagct agtgcggaag atgatgttaa agctatcaag gatcttttag 3061 atcaaactaa taatctctta cataaactaa aaatatttca tattagtcag tcagaagata 3121 aggcaaatat tttagacaag gatgagcatt tttatctagt atttgaggag tgctactttg 3181 agctagcgaa tatagtgcct ctttataaca aaattagaaa ctatataact caaaagccat 3241 atagtgatga gaaatttaag ctcaattttg agaactcgac tttggctaat ggttgggata 3301 aaaataaaga gcctgacaat acggcaattt tatttatcaa agatgataaa tattatctgg 3361 gtgtgatgaa taagaaaaat aacaaaatat ttgatgataa agctatcaaa gaaaataaag 3421 gcgagggtta taaaaaaatt gtttataaac ttttacctgg cgcaaataaa atgttaccta 3481 aggttttctt ttctgctaaa tctataaaat tttataatcc tagtgaagat atacttagaa 3541 taagaaatca ttccacacat acaaaaaatg gtagtcctca aaaaggatat gaaaaatttg 3601 agtttaatat tgaagattgc cgaaaattta tagattttta taaacagtct ataagtaagc 3661 atccggagtg gaaagatttt ggatttagat tttctgatac tcaaagatat aattctatag 3721 atgaatttta tagagaagtt gaaaatcaag gctacaaact aacttttgaa aatatatcag 3781 agagctatat tgatagcgta gttaatcagg gtaaattgta cctattccaa atctataata 3841 aagatttttc agcttatagc aaagggcgac caaatctaca tactttatat tggaaagcgc 3901 tgtttgatga gagaaatctt caagatgtgg tttataagct aaatggtgag gcagagcttt 3961 tttatcgtaa acaatcaata cctaaaaaaa tcactcaccc agctaaagag gcaatagcta 4021 ataaaaacaa agataatcct aaaaaagaga gtgtttttga atatgattta atcaaagata 4081 aacgctttac tgaagataag tttttctttc actgtcctat tacaatcaat tttaaatcta 4141 gtggagctaa taagtttaat gatgaaatca atttattgct aaaagaaaaa gcaaatgatg 4201 ttcatatatt aagtatagat agaggtgaaa gacatttagc ttactatact ttggtagatg 4261 gtaaaggcaa tatcatcaaa caagatactt tcaacatcat tggtaatgat agaatgaaaa 4321 caaactacca tgataagctt gctgcaatag agaaagatag ggattcagct aggaaagact 4381 ggaaaaagat aaataacatc aaagagatga aagagggcta tctatctcag gtagttcatg 4441 aaatagctaa gctagttata gagtataatg ctattgtggt ttttgaggat ttaaattttg 4501 gatttaaaag agggcgtttc aaggtagaga agcaggtcta tcaaaagtta gaaaaaatgc 4561 taattgagaa actaaactat ctagttttca aagataatga gtttgataaa actgggggag 4621 tgcttagagc ttatcagcta acagcacctt ttgagacttt taaaaagatg ggtaaacaaa 4681 caggtattat ctactatgta ccagctggtt ttacttcaaa aatttgtcct gtaactggtt 4741 ttgtaaatca gttatatcct aagtatgaaa gtgtcagcaa atctcaagag ttctttagta 4801 agtttgacaa gatttgttat aaccttgata agggctattt tgagtttagt tttgattata 4861 aaaactttgg tgacaaggct gccaaaggca agtggactat agctagcttt gggagtagat 4921 tgattaactt tagaaattca gataaaaatc ataattggga tactcgagaa gtttatccaa 4981 ctaaagagtt ggagaaattg ctaaaagatt attctatcga atatgggcat ggcgaatgta 5041 tcaaagcagc tatttgcggt gagagcgaca aaaagttttt tgctaagcta actagtgtcc 5101 taaatactat cttacaaatg cgtaactcaa aaacaggtac tgagttagat tatctaattt 5161 caccagtagc agatgtaaat ggcaatttct ttgattcgcg acaggcgcca aaaaatatgc 5221 ctcaagatgc tgatgccaat ggtgcttatc atattgggct aaaaggtctg atgctactag 5281 gtaggatcaa aaataatcaa gagggcaaaa aactcaattt ggttatcaaa aatgaagagt 5341 attttgagtt cgtgcagaat aggaataact aattgacagc tagctcagtc ctaggtataa 5401 tgctagcgct gatttaggca aaaacgggtc taagaacttt aaataatttc tactgttgta 5461 gatgagaagt catttaataa ggccactgtt aaaagtctaa gaactttaaa taatttctac 5521 tgttgtagat gctactattc ctgtgccttc agataattca gtctaagaac tttaaataat 5581 ttctactgtt gtagatgtct agagcctttt gtattagtag ccggtctaag aactttaaat 5641 aatttctact gttgtagatt agcgatttat gaaggtcatt tttttgtcta gctttaatgc 5701 ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat 5761 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg 5821 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat 5881 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca 5941 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc 6001 gactacgcga tcatggcgac cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg 6061 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat 6121 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg 6181 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg 6241 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat 6301 aagggagagc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg 6361 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta 6421 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg 6481 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc 6541 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg 6601 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc 6661 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc 6721 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc 6781 ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca 6841 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg 6901 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta 6961 acggattcac cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc 7021 aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc 7081 gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga 7141 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg 7201 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg 7261 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcccctacg tgctgctgaa 7321 gttgcccgca acagagagtg gaaccaaccg gtgataccac gatactatga ctgagagtca 7381 acgccatgag cggcctcatt tcttattctg agttacaaca gtccgcaccg ctgtccggta 7441 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta 7501 tcatgcaact cgtaggacag gtgccggcag cgcccaacag tcccccggcc acggggcctg 7561 ccaccatacc cacgccgaaa caagcgccct gcaccattat gttccggatc tgcatcgcag 7621 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc taaccgtttt 7681 tatcaggctc tgggaggcag aataaatgat catatcgtca attattacct ccacggggag 7741 agcctgagca aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt 7801 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc 7861 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt 7921 caggcgtagc accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc 7981 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat 8041 cacagacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat 8101 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat 8161 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc 8221 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta 8281 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct 8341 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca 8401 ttgccatacg //