LOCUS Exported 8542 bp ds-DNA circular SYN 11-SEP-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 8542) AUTHORS Yiping Qi Lab TITLE Direct Submission JOURNAL Exported Nov 11, 2019 from SnapGene Viewer 4.0.7 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8542 /organism="recombinant plasmid" /mol_type="other DNA" promoter 1..138 /label=Spn promoter /note="ApEinfo_fwdcolor=pink ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink dnas_title=Spn promoter vntifkey=30" CDS 139..1149 /gene="Streptomycin/Spectinomycin adenyltransferase" /label=SpnR /note="ApEinfo_fwdcolor=pink ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink dnas_title=SpnR invitrogen=2750000 vntifkey=4" rep_origin 1216..1889 /gene="ori" /label=pUC origin /note="ApEinfo_fwdcolor=pink ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink dnas_title=pUC origin invitrogen=2530000 vntifkey=33" terminator 2319..2362 /label=rrnB T1 terminatior terminator /label=rrnB T1 terminatior /note="ApEinfo_fwdcolor=pink ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink dnas_title=rrnB T1 terminatior vntifkey=43" primer_bind 2429..2444 /label=M13 (-20) forward primer misc_recomb 2461..2560 /label=attL1 /note="ApEinfo_fwdcolor=#00ff40 ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink dnas_title=attL1 invitrogen=2670000 vntifkey=86" gene 2579..2629 /label=N-terminal NLS /note="N-terminal NLS ApEinfo_fwdcolor=pink ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink" misc_feature 2585..2605 /label=SV40 NLS /note="SV40 NLS ApEinfo_fwdcolor=#ff0000 ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=#ff0000" CDS 2630..6526 /codon_start=1 /label=dFnCas12a /translation="SIYQEFVNKYSLSKTLRFELIPQGKTLENIKARGLILDDEKRAKD YKKAKQIIDKYHQFFIEEILSSVCISEDLLQNYSDVYFKLKKSDDDNLQKDFKSAKDTI KKQISEYIKDSEKFKNLFNQNLIDAKKGQESDLILWLKQSKDNGIELFKANSDITDIDE ALEIIKSFKGWTTYFKGFHENRKNVYSSNDIPTSIIYRIVDDNLPKFLENKAKYESLKD KAPEAINYEQIKKDLAEELTFDIDYKTSEVNQRVFSLDEVFEIANFNNYLNQSGITKFN TIIGGKFVNGENTKRKGINEYINLYSQQINDKTLKKYKMSVLFKQILSDTESKSFVIDK LEDDSDVVTTMQSFYEQIAAFKTVEEKSIKETLSLLFDDLKAQKLDLSKIYFKNDKSLT DLSQQVFDDYSVIGTAVLEYITQQIAPKNLDNPSKKEQELIAKKTEKAKYLSLETIKLA LEEFNKHRDIDKQCRFEEILANFAAIPMIFDEIAQNKDNLAQISIKYQNQGKKDLLQAS AEDDVKAIKDLLDQTNNLLHKLKIFHISQSEDKANILDKDEHFYLVFEECYFELANIVP LYNKIRNYITQKPYSDEKFKLNFENSTLANGWDKNKEPDNTAILFIKDDKYYLGVMNKK NNKIFDDKAIKENKGEGYKKIVYKLLPGANKMLPKVFFSAKSIKFYNPSEDILRIRNHS THTKNGSPQKGYEKFEFNIEDCRKFIDFYKQSISKHPEWKDFGFRFSDTQRYNSIDEFY REVENQGYKLTFENISESYIDSVVNQGKLYLFQIYNKDFSAYSKGRPNLHTLYWKALFD ERNLQDVVYKLNGEAELFYRKQSIPKKITHPAKEAIANKNKDNPKKESVFEYDLIKDKR FTEDKFFFHCPITINFKSSGANKFNDEINLLLKEKANDVHILSIARGERHLAYYTLVDG KGNIIKQDTFNIIGNDRMKTNYHDKLAAIEKDRDSARKDWKKINNIKEMKEGYLSQVVH EIAKLVIEYNAIVVFEDLNFGFKRGRFKVEKQVYQKLEKMLIEKLNYLVFKDNEFDKTG GVLRAYQLTAPFETFKKMGKQTGIIYYVPAGFTSKICPVTGFVNQLYPKYESVSKSQEF FSKFDKICYNLDKGYFEFSFDYKNFGDKAAKGKWTIASFGSRLINFRNSDKNHNWDTRE VYPTKELEKLLKDYSIEYGHGECIKAAICGESDKKFFAKLTSVLNTILQMRNSKTGTEL DYLISPVADVNGNFFDSRQAPKNMPQDADANGAYHIGLKGLMLLGRIKNNQEGKKLNLV IKNEEYFEFVQNRNN" misc_feature 2630..6526 /label=dFnCas12a /label=FnCpf1 /note="ApEinfo_fwdcolor=#00ff46 ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=green" misc_feature 5362..5381 /label=FnCas12aD917A-R misc_feature 5375..5377 /label=D917A misc_feature 5382..5401 /label=FnCas12aD917A-F misc_feature 6527..6574 /label=Nucleoplasmin NLS /note="Nucleoplasmin NLS ApEinfo_fwdcolor=#ff0000 ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=#ff0000" misc_feature 6575..6616 /label=5XGS linker /note="5XGS linker" CDS 6617..6769 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6785..6937 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6959..7111 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 7127..7279 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 7301..7453 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 7469..7621 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 7658..7807 /codon_start=1 /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 7820..7969 /codon_start=1 /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" misc_feature 7985..7989 /label=AatII /note="AatII" misc_feature 7990..8267 /label=NOS term /note="ApEinfo_fwdcolor=#f5010a ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=green dnas_title=NOS term vntifkey=21" misc_recomb 8289..8412 /label=attR5 /note="ApEinfo_fwdcolor=#00ff00 ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_revcolor=pink dnas_title=attR5 invitrogen=2703000 vntifkey=86" primer_bind complement(8453..8469) /label=M13 reverse primer promoter 8541..8542 /label=Spn promoter(1) promoter /label=Spn promoter(1) /note="ApEinfo_fwdcolor=pink ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ApEinfo_label=Spn promoter ApEinfo_revcolor=pink dnas_title=Spn promoter vntifkey=30" ORIGIN 1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc 61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag 121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc 181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag 241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta 301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacatcatg 361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag 421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc 481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca 541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag 601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat 661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc 721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat 781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat 841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc 901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa 961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag 1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg 1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc 1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc 1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc 2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa 2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc 2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat 2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt 2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt 2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc 2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattcgc ccttcaccat 2581 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggctt cgatctatca 2641 agaattcgtt aataagtatt cgctttctaa aacactgaga ttcgaactca tacctcaagg 2701 aaagacactc gagaacataa aggctagggg cctcatattg gatgatgaaa agagagccaa 2761 ggactataaa aaagccaaac agataatcga caagtaccac caattcttca tagaagaaat 2821 cctttccagc gtctgcatta gcgaggattt gttgcaaaat tacagcgatg tctacttcaa 2881 gcttaagaag agcgatgacg acaatctcca gaaagacttc aaatcagcga aggacacaat 2941 taagaagcag atcagcgagt atatcaaaga tagtgaaaaa ttcaagaacc ttttcaatca 3001 gaacctgata gacgcaaaaa aaggacaaga aagcgacctc attctttggt tgaagcagtc 3061 taaggacaac gggattgagc tttttaaggc gaatagcgat ataaccgaca tcgacgaggc 3121 gcttgagatc ataaagtcgt ttaagggatg gacaacatac ttcaaaggct tccatgagaa 3181 tcgcaaaaac gtctactcca gcaacgacat tccaacgtcg attatataca gaattgttga 3241 tgataacctc cctaaattcc tcgaaaataa ggcaaaatat gaaagcctta aagataaagc 3301 gcctgaagca atcaattatg aacaaatcaa aaaggatctt gctgaagaat tgacgtttga 3361 tatagactac aagacgtcag aagttaacca gagggtgttt tcactcgacg aggtgtttga 3421 gattgctaat ttcaacaact acctcaatca gagtgggatc acgaagttca acactattat 3481 aggtggtaag ttcgtgaatg gtgagaacac taaaagaaaa gggattaacg aatatataaa 3541 cctttatagt caacagatca acgacaagac tttgaagaaa tataagatga gcgtcctctt 3601 caagcagata ctcagtgaca cggaatccaa gagctttgtg atcgacaagc tcgaagatga 3661 ctcggatgtg gtcactacca tgcaatcctt ctacgagcaa attgccgctt tcaaaactgt 3721 ggaggagaag agtataaagg agacactgtc tctgcttttt gacgatctta aggcccagaa 3781 attggatctt tccaaaatat atttcaagaa tgataagtca cttacggacc tttcccaaca 3841 agtttttgac gattattcag ttattggtac ggcggttctt gagtacatta cgcagcagat 3901 agcccccaag aatctggaca acccctctaa gaaagaacag gagttgatag cgaagaagac 3961 agagaaggcg aaatacctct cgctggagac cataaaattg gcactggaag aatttaacaa 4021 gcatcgcgac atagataaac agtgccgctt cgaggaaatt ttggcaaatt ttgcagccat 4081 tccaatgatt ttcgacgaga tagcgcaaaa caaggataat ttggcacaaa tttcaataaa 4141 atatcaaaac cagggcaaga aggacctctt gcaggcttcg gcagaggatg atgttaaggc 4201 tattaaagac ttgttggacc aaacgaacaa tctgttgcac aagttgaaaa ttttccacat 4261 tagtcaaagc gaggataaag caaacatatt ggacaaagac gagcactttt atctggtgtt 4321 tgaggagtgc tacttcgagc ttgctaatat tgtcccactt tataataaaa taagaaacta 4381 cattacgcaa aagccatatt cagatgaaaa gtttaagctc aatttcgaaa atagtactct 4441 tgccaacggc tgggacaaga ataaggagcc agataatacc gccatacttt ttatcaaaga 4501 tgataaatat tatcttgggg tgatgaataa gaagaataat aagatcttcg atgataaagc 4561 gataaaggaa aataagggtg aaggctataa aaaaattgtt tacaaactgt tgccgggagc 4621 aaataaaatg ctccccaagg tttttttttc ggcaaagagc attaaatttt acaatccttc 4681 agaagacatt ctgcgcataa gaaatcattc gacacacact aaaaatggtt cgccacaaaa 4741 gggctacgag aaatttgaat tcaacattga ggactgtcgg aagttcattg atttctacaa 4801 gcagtccatc tccaagcacc cggagtggaa agattttggg tttcggtttt ccgacacgca 4861 gagatacaac agcattgatg aattttatag agaggtcgag aatcaaggtt ataagcttac 4921 ctttgaaaac atttctgaat catacattga ttcagtggtc aatcagggca aactctatct 4981 ttttcaaata tacaacaagg actttagtgc ttatagtaaa gggcggccca atttgcatac 5041 tctctattgg aaagcgctgt ttgatgagcg gaaccttcaa gacgtcgtgt ataagctcaa 5101 cggggaagcc gagctctttt accgcaagca gtccataccg aaaaaaataa cacaccctgc 5161 caaagaagcc atcgccaaca agaataaaga caatcctaaa aaagagtccg tcttcgaata 5221 tgatcttatt aaggacaaga ggtttacaga agataaattt ttcttccatt gtcccataac 5281 tatcaatttc aaaagctctg gcgcgaacaa atttaacgac gaaatcaatc tcttgttgaa 5341 agaaaaagcc aacgatgtgc acattctgtc gatcgccagg ggagagcgcc acttggcata 5401 ctacaccctt gttgatggga aaggaaatat tattaaacag gacacattta atatcatcgg 5461 caacgatcgc atgaagacca actatcatga caaactggca gcaattgaaa aggaccgcga 5521 ctcagcgaga aaagactgga agaagatcaa taatatcaaa gaaatgaaag agggttattt 5581 gtctcaagtg gtccatgaga tcgcgaagtt ggtcattgaa tataatgcca tagtggtctt 5641 cgaagatctg aattttggat ttaagcgcgg caggttcaaa gtcgaaaaac aggtctacca 5701 aaagttggaa aagatgctca tcgaaaagct gaattacctt gtcttcaaag ataacgaatt 5761 cgataaaacc gggggggtct tgagggccta ccaactgact gcaccctttg agacttttaa 5821 aaagatgggt aaacagacag gaataattta ctatgttcct gccggtttca ctagcaagat 5881 ttgccccgtt accggattcg tgaatcaact ctatcccaaa tacgaatccg tgagcaagag 5941 tcaggaattc ttctccaaat ttgataaaat atgctataat ctcgacaaag gttatttcga 6001 gttctcgttc gactataaga acttcgggga taaggctgcc aagggaaagt ggactatagc 6061 aagctttggt agtcgcctta taaattttag gaacagcgac aagaatcaca actgggacac 6121 tcgggaagtc tacccaacaa aagaactgga gaaactcttg aaggattata gtatcgagta 6181 tgggcatggg gagtgtatca aggcagcgat ttgtggagag tccgacaaaa agttttttgc 6241 taaactcacc tcggtgctca acactatcct ccagatgaga aattcaaaaa cagggacaga 6301 gctcgattac ctcattagcc ccgttgccga cgtcaatgga aactttttcg actcaagaca 6361 ggctccaaaa aacatgccgc aagatgcgga cgcgaatggg gcctatcaca taggcctgaa 6421 agggcttatg ctccttggga gaattaaaaa taaccaagaa ggcaaaaaac tcaacctcgt 6481 cattaagaac gaagaatact tcgaatttgt tcagaacagg aataacaagc gtcctgctgc 6541 caccaaaaag gccggacagg ctaagaaaaa gaagggagac ggctctggat cggggtcggg 6601 ttctggctca gtcgacttgc ttgatccggg gacaccaatg gacgcggacc tggtggcttc 6661 atcgaccgtg gtttgggaac aggacgccga tccattcgcc gggaccgccg atgactttcc 6721 tgcttttaat gaggaagagt tggcttggct gatggaactc ctgccgcagg gcggctcagg 6781 gggtctcctt gaccccggca cccccatgga cgctgacctc gttgcaagtt cgacggttgt 6841 ttgggagcaa gatgcagatc cgtttgcggg tacagctgat gactttccag ccttcaacga 6901 agaggagctg gcatggctta tggagcttct gcctcaggcc agaggagggt cgggtgggct 6961 cttggatcct gggacgccga tggatgccga ccttgtcgcg tcctctactg tggtttggga 7021 gcaggacgcc gacccgttcg ctggtactgc tgatgacttt ccggccttta atgaggagga 7081 actcgcttgg ctcatggagt tgttgccaca agggggtagt ggcggtcttc tggacccggg 7141 aacgcctatg gatgctgact tggtggcatc gagcaccgtc gtctgggaac aggatgcgga 7201 cccgtttgct ggtaccgctg atgattttcc ggcatttaac gaggaagaat tggcgtggct 7261 catggaattg cttccccagg cgagaggggg ttccggtggc ttgctggatc cgggaacgcc 7321 tatggatgcc gatctcgtgg cgtcgtcaac cgtggtctgg gagcaagacg ctgacccgtt 7381 tgccggcaca gcagatgatt ttccagcttt caatgaggaa gaactggcat ggttgatgga 7441 attgcttcca cagggaggga gcggaggcct cctcgatcct ggaacaccta tggacgcaga 7501 cctggttgcc tcttcaactg tcgtgtggga acaagatgca gacccgttcg caggtacagc 7561 tgatgacttc cctgccttta atgaggagga actcgcgtgg ctcatggaac ttttgcccca 7621 agcacgcggt ggctctgggg gtggcggatc gggaggagat gcgctggacg atttcgactt 7681 ggacatgttg ggctcagacg ctttggacga ctttgatctc gacatgcttg ggtccgacgc 7741 attggatgat ttcgaccttg acatgcttgg ttccgacgca ctcgatgact tcgatcttga 7801 tatgctcgcc cgcggatctg acgctctcga cgacttcgat cttgacatgt tgggctcaga 7861 tgcgctcgac gactttgatc tcgatatgtt gggtagcgac gcactcgatg actttgactt 7921 ggatatgctg gggagcgacg ccttggacga tttcgatctg gacatgctgt aaaagctttg 7981 ataggacgtc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg 8041 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta 8101 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat 8161 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg 8221 cggtgtcatc tatgttacta gatcgggaat tgatcccccc tcgacagctt ccggaaaggg 8281 cgaattcgca actttgtata caaaagttga acgagaaacg taaaatgata taaatatcaa 8341 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat 8401 ccagtcacta tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc 8461 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa 8521 aaatatatca tcatgcctcc tc //