LOCUS AID'-BE4max 8763 bp ds-DNA circular 04-AUG-2021 DEFINITION . FEATURES Location/Qualifiers misc_feature 1..30 /label="SV40 NLS" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 promoter 61..364 /label="CMV enhancer" /ApEinfo_revcolor=#c7b0e3 /ApEinfo_fwdcolor=#c7b0e3 promoter 365..568 /label="CMV promoter" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 misc_feature 582..603 /label="overlap gibson" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd primer_bind 610..629 /label="T7" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 642..663 /label="Primer " /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 642..1184 /label="AIDC12(1-181) " /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e CDS 642..5951 /label="Translation 642-5951" primer complement(786..803) /label="AIDC12-E58A reverse" /note="sequence: GCCGTTTTTGTTACGCAG" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 primer 804..824 /label="AIDC12-E58A forward" /note="sequence: TGCCATGTGGCACTGCTGTTT" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature 1164..1184 /label="primer " /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 1185..1280 /label="linker" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 misc_feature 1195..1224 /label="overlap gibson" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc primer complement(1198..1215) /label="XmaI for BE4max Reverse" /note="sequence: tgccagaggAtcctccgc" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 primer 1216..1240 /label="XmaI for BE4max Forward" /note="sequence: gCGAGACACCcgggACAAGCGAGTC" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 misc_feature 1229..1251 /label="overlap gibson" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 misc_feature 1281..5381 /label="nCas9" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 misc_feature 5382..5951 /label="2X UGI" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc misc_feature 5952..6002 /label="NLS" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 CDS 5952..6030 /label="Translation 5952-6030" primer_bind complement(6348..6368) /label="M13-rev" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_binding complement(6374..6396) /label="LacO" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc promoter complement(6400..6430) /label="lac promoter" /ApEinfo_revcolor=#c7b0e3 /ApEinfo_fwdcolor=#c7b0e3 misc_feature complement(6401..6430) /label="lac promoter" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 misc_feature 7513..8373 /label="Amp resistance " /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature complement(8415..8443) /label="amp promoter" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc 601 cgcggccgct aatacgactc actataggga gagccgccac cATGGATAGC CTGCTGATGA 661 ACCGTCGTgA ATTTCTGTAT CAGTTTAAAA ACGTGCGTTG GGCGAAAGGC CGTCGTGAAA 721 CCTATCTGTG CTATGTGGTG AAACGTCGTG ATAGCGCGAC CAGCTTTAGC CTGGATTTTG 781 GCTATCTGCG TAACAAAAAC GGCTGCCATG TGGAACTGCT GTTTCTGCGT TATATTAGCG 841 ATTGGGATCT GGATCCGGGC CGTTGCTATC GTGTGACCTG GTTTAtCAGC TGGAGCCCGT 901 GCTATGATTG CGCGCGTCAT GTGGCGGATT TTCTGCGTGG CAACCCGAAC CTGAGCCTGC 961 GTATTTTTAC CGCGCGTCTG TATTTTTGCG AAgCcGgcaG gCGtGAACCG GAAGGCCTGC 1021 GTCGTCTGCA TCGTGCGGGC GTGCAGATTG CGATTATGAC CTTTAAAGAT TATTTTTATT 1081 GCTGGAACAC CTTTGTGGAA AACCATGgAC GTACCTTTAA AGCGTGGGAA GGCCTGCATG 1141 AAAACAGCGT GCGTCTGAGC CGTCAGCTGC GTCGTATTCT GCTGtctggc ggatctagcg 1201 gaggatcctc tggcagcgag acacccggga caagcgagtc agcaacacca gagagcagtg 1261 gcggcagcag cggcggcagc gacaagaagt acagcatcgg cctggccatc ggcaccaact 1321 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc 1381 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca 1441 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac 1501 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg 1561 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc 1621 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca 1681 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct 1741 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga 1801 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc 1861 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca 1921 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga 1981 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca 2041 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc 2101 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga 2161 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg 2221 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc 2281 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga 2341 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt 2401 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca 2461 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc 2521 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg 2581 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc 2641 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc 2701 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga 2761 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt 2821 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga 2881 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga 2941 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct 3001 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc 3061 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca 3121 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac 3181 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga 3241 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt 3301 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc 3361 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg 3421 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga 3481 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca 3541 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga 3601 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga 3661 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact 3721 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg 3781 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca 3841 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg 3901 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga 3961 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg 4021 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc 4081 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag 4141 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag 4201 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa 4261 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg 4321 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca 4381 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg 4441 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg 4501 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg 4561 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga 4621 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg 4681 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca 4741 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg 4801 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga 4861 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg 4921 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact 4981 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga 5041 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg 5101 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca 5161 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc 5221 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga 5281 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc 5341 tgtacgagac acggatcgac ctgtctcagc tgggaggtga cagcggcggg agcggcggga 5401 gcgaggggag cactaatctg agcgacatca ttgagaagaa gactgggaaa cagctggtca 5461 ttcaggagtc catcctgatg ctgcctgagg aggtggagga agtgatcggc aacaagccag 5521 agtctgacat cctggtgcac accgcctacg acgagtccac agatgagaat gtgatgctgc 5581 tgacctctga cgcccccgag tataagcctt gggccctggt catccaggat tctaacggcg 5641 agaataagat caagatgctg agcggaggat ccggaggatc tggaggcagc accaacctgt 5701 ctgacatcat cgagaaggag acaggcaagc agctggtcat ccaggagagc atcctgatgc 5761 tgcccgaaga agtcgaagaa gtgatcggaa acaagcctga gagcgatatc ctggtccata 5821 ccgcctacga cgagagtacc gacgaaaatg tgatgctgct gacatccgac gccccagagt 5881 ataagccctg ggctctggtc atccaggatt ccaacggaga gaacaaaatc aaaatgctgt 5941 ctggcggctc aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag 6001 tctaaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt 6061 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 6121 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 6181 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 6241 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 6301 cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 6361 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 6421 aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 6481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 6541 cgcgggaaga ggcggtttgc gtattgggcg cacttccgct tcctcgctca ctgactcgct 6601 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 6661 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 6721 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 6781 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 6841 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 6901 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 6961 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 7021 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 7081 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 7141 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 7201 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 7261 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 7321 gcgcagaaaa aaaggatctc aagaaaatcc tttgatcttt tctacggggt ctgacactca 7381 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7441 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 7501 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 7561 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 7621 accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt 7681 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 7741 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 7801 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 7861 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 7921 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7981 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 8041 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 8101 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 8161 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 8221 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 8281 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8341 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 8401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 8461 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 8521 agatcgatct cccgatcccc tagggtcgac tctcagtaca atctgctctg atgccgcata 8581 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa 8641 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt 8701 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt 8761 gac //