LOCUS Exported 8029 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 8029) 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..8029 /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 2412..2435 /label=M13-F1 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" CDS 2579..2629 /codon_start=1 /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" /translation="MAPKKKRKVGIHGVPAA" 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" gene 2630..6013 /label=AaCas12b (C2C1; rice codon optimized) primer_bind 3025..3045 /label=292-seq1 primer_bind 3632..3652 /label=292-seq2 primer_bind 4196..4215 /label=292-seq3 misc_feature 4334..4336 /label=D570A primer_bind 4805..4824 /label=292-seq4 primer_bind 5421..5440 /label=292-seq5 CDS 6014..6061 /codon_start=1 /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" /translation="KRPAATKKAGQAKKKK" misc_feature 6062..6103 /label=5XGS linker /note="5XGS linker" CDS 6104..6256 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6272..6424 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6446..6598 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6614..6766 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6788..6940 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 6956..7108 /codon_start=1 /label=TAL /translation="LLDPGTPMDADLVASSTVVWEQDADPFAGTADDFPAFNEEELAWL MELLPQ" CDS 7145..7294 /codon_start=1 /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 7307..7456 /codon_start=1 /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" misc_feature 7472..7476 /label=AatII /note="AatII" misc_feature 7477..7754 /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 7776..7899 /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(7940..7956) /label=M13 reverse primer promoter 8028..8029 /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 cctgcggctg ccgtcaagtc 2641 catgaaggtc aagttgcgcc tggataacat gccagagatc agagccggac tttggaaact 2701 tcacaccgag gttaatgcgg gtgtgcggta ctatacggaa tggcttagcc ttttgaggca 2761 agaaaatctt tatcggagga gtcccaatgg cgatggagaa caagaatgct ataaaactgc 2821 tgaggaatgc aaggctgaac tccttgagag actcagagcc cgccaagttg agaatgggca 2881 ctgcggccct gctgggagtg atgacgaact gctgcaattg gcacggcaac tttatgaact 2941 tctggtccca caagcaatcg gggctaaagg tgatgcgcag caaatcgcaa ggaagtttct 3001 tagtcccctt gccgacaagg atgccgtggg tggtttggga atagcaaaag caggaaataa 3061 gcctaggtgg gttcggatga gggaggctgg agagccaggt tgggaagagg aaaaggctaa 3121 agccgaggcg agaaagagta cggatagaac cgccgatgtt cttcgcgctc ttgcagactt 3181 cggtcttaaa cctcttatga gagtctacac agactcagac atgtccagcg tgcagtggaa 3241 accacttcgc aaaggacaag cggtcagaac ctgggataga gacatgttcc aacaagcgat 3301 cgaaagaatg atgagttggg aatcgtggaa tcagcgcgtt ggagaagcgt acgcaaagct 3361 cgtggaacaa aagtcgaggt ttgaacagaa aaattttgtg ggacaagaac atcttgtcca 3421 acttgtcaat caacttcaac aagacatgaa ggaagcatca cacggcctgg agtcgaaaga 3481 acaaactgcg cattacttga ctgggagagc gctgagaggg agcgacaaag tttttgagaa 3541 gtgggaaaaa ctcgatcctg atgccccatt tgacctctat gataccgaaa tcaagaatgt 3601 tcaacggagg aatactcgca ggttcggatc tcatgatctg tttgcgaagc tcgcggaacc 3661 taaatatcag gcgctctgga gagaggacgc ttctttcctc acgaggtatg cggtttacaa 3721 tagcattgtc agaaaactga atcacgctaa aatgtttgcg acttttactc ttccggatgc 3781 taccgcccac ccgatctgga cgcggtttga caaactcggc ggcaacctgc accagtacac 3841 tttcttgttt aacgaatttg gcgagggcag gcacgccatt cggtttcaga agctgttgac 3901 ggttgaggat ggcgttgcta aagaggtcga cgacgtcacg gttccgattt ctatgtccgc 3961 gcagctggat gacctcttgc ctcgggaccc acacgagctc gttgcactct acttccagga 4021 ctacggtgca gaacaacatc tggctggaga gtttggcggc gcgaaaattc aataccgccg 4081 cgatcaattg aaccacctgc acgccagaag aggcgccaga gatgtctacc ttaatctgag 4141 cgtccgcgtt cagtcacaat ccgaagccag gggagaaagg cgccctccgt atgcagcggt 4201 cttcaggctt gttggcgata accaccgcgc gtttgttcac tttgataaat tgtcagatta 4261 cctcgcagaa cacccagacg atggtaagct ggggtcggaa ggtttgctct ctgggctcag 4321 agtcatgtca gttgccttgg gtcttaggac ttccgcgagc atatctgtct tccgcgtcgc 4381 aagaaaggac gaattgaagc cgaacagtga aggccgggtc cctttttgct tcccgatcga 4441 agggaacgaa aacctcgttg ctgtccacga gcggagccaa ctgttgaagc ttcccggtga 4501 aacggaatcg aaagatctga gagcgatcag agaagagcgc caaaggacgc ttagacagct 4561 ccggacgcaa cttgcatact tgcgccttct ggttcgctgc ggtagtgaag acgttggaag 4621 aagagagagg tcatgggcta aactcataga gcaacctatg gatgctaatc aaatgacgcc 4681 tgattggaga gaagcattcg aagacgaact tcagaaactg aaatcccttt acgggatatg 4741 cggcgatcgc gagtggacag aagcagtgta tgagtctgtg aggcgcgtgt ggcggcatat 4801 gggtaaacag gtgcgcgatt ggagaaaaga cgttaggagc ggggaaagac ctaagatacg 4861 gggatatcag aaagacgttg tcgggggaaa tagcattgaa cagattgaat atttggagcg 4921 ccaatataag ttcctcaaat cctggtcttt cttcggcaaa gtgtcaggcc aggtgatacg 4981 cgcggaaaag ggatcgcgct ttgcaataac tctgagagaa catattgatc atgccaaaga 5041 agatcggttg aagaaactcg ccgatagaat catcatggag gcgcttggtt atgtctacgc 5101 cttggacgat gaacggggaa agggaaagtg ggtcgccaag tatccacctt gccaactcat 5161 tctcctcgaa gaactttccg aataccagtt taacaacgat cggccgccat cagagaataa 5221 tcaactgatg cagtggtccc atcgcggtgt gtttcaagag ttgctcaatc aggcccaagt 5281 ccatgatctg cttgttggca caatgtatgc agccttttcc tcccggtttg atgcaagaac 5341 aggggctcct ggcatacgct gtagacgggt cccggcgagg tgcgcccgcg aacaaaaccc 5401 tgaaccgttc ccctggtggt tgaacaagtt cgttgcggag cacaagctgg acgggtgtcc 5461 tctgcgggcc gacgatctta ttcccaccgg ggaaggggaa ttctttgtga gccctttctc 5521 ggcggaggaa ggggattttc accaaataca tgcagatctt aatgccgcac aaaatttgca 5581 gaggagactg tggtcagact ttgatattag tcagatacgc ctccgctgtg actggggaga 5641 ggtcgatggc gagcctgtgt tgataccaag aacgaccgga aagaggacag ccgattcgta 5701 tggaaacaag gttttttaca cgaagacggg cgttacttac tacgaaagag aaagagggaa 5761 gaagagaagg aaagtctttg cccaagaaga attgagcgag gaagaagccg agctcttggt 5821 cgaagcggac gaggcacggg aaaagtctgt cgtcctcatg agggaccctt ccggaattat 5881 taaccgggga gattggacgc ggcagaaaga gttttggtcc atggttaatc aacgcataga 5941 aggctacctt gtcaagcaaa taagaagtcg cgtgagattg caggagagtg catgtgagaa 6001 cactggggac ataaagcgtc ctgctgccac caaaaaggcc ggacaggcta agaaaaagaa 6061 gggagacggc tctggatcgg ggtcgggttc tggctcagtc gacttgcttg atccggggac 6121 accaatggac gcggacctgg tggcttcatc gaccgtggtt tgggaacagg acgccgatcc 6181 attcgccggg accgccgatg actttcctgc ttttaatgag gaagagttgg cttggctgat 6241 ggaactcctg ccgcagggcg gctcaggggg tctccttgac cccggcaccc ccatggacgc 6301 tgacctcgtt gcaagttcga cggttgtttg ggagcaagat gcagatccgt ttgcgggtac 6361 agctgatgac tttccagcct tcaacgaaga ggagctggca tggcttatgg agcttctgcc 6421 tcaggccaga ggagggtcgg gtgggctctt ggatcctggg acgccgatgg atgccgacct 6481 tgtcgcgtcc tctactgtgg tttgggagca ggacgccgac ccgttcgctg gtactgctga 6541 tgactttccg gcctttaatg aggaggaact cgcttggctc atggagttgt tgccacaagg 6601 gggtagtggc ggtcttctgg acccgggaac gcctatggat gctgacttgg tggcatcgag 6661 caccgtcgtc tgggaacagg atgcggaccc gtttgctggt accgctgatg attttccggc 6721 atttaacgag gaagaattgg cgtggctcat ggaattgctt ccccaggcga gagggggttc 6781 cggtggcttg ctggatccgg gaacgcctat ggatgccgat ctcgtggcgt cgtcaaccgt 6841 ggtctgggag caagacgctg acccgtttgc cggcacagca gatgattttc cagctttcaa 6901 tgaggaagaa ctggcatggt tgatggaatt gcttccacag ggagggagcg gaggcctcct 6961 cgatcctgga acacctatgg acgcagacct ggttgcctct tcaactgtcg tgtgggaaca 7021 agatgcagac ccgttcgcag gtacagctga tgacttccct gcctttaatg aggaggaact 7081 cgcgtggctc atggaacttt tgccccaagc acgcggtggc tctgggggtg gcggatcggg 7141 aggagatgcg ctggacgatt tcgacttgga catgttgggc tcagacgctt tggacgactt 7201 tgatctcgac atgcttgggt ccgacgcatt ggatgatttc gaccttgaca tgcttggttc 7261 cgacgcactc gatgacttcg atcttgatat gctcgcccgc ggatctgacg ctctcgacga 7321 cttcgatctt gacatgttgg gctcagatgc gctcgacgac tttgatctcg atatgttggg 7381 tagcgacgca ctcgatgact ttgacttgga tatgctgggg agcgacgcct tggacgattt 7441 cgatctggac atgctgtaaa agctttgata ggacgtccga tcgttcaaac atttggcaat 7501 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt 7561 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg 7621 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc 7681 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattga 7741 tcccccctcg acagcttccg gaaagggcga attcgcaact ttgtatacaa aagttgaacg 7801 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac 7861 tacataatac tgtaaaacac aacatatcca gtcactatgc catccagctg atatccccta 7921 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa 7981 tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctc //