LOCUS sgRNA_eGFP_314 10394 bp ds-DNA circular 12-OCT-2017 DEFINITION . FEATURES Location/Qualifiers promoter 1380..1583 /label="CMV promoter" misc_feature 763..963 /label="g-block" rep_origin 6034..6169 /label="SV40 ori" rep_origin 9796..10384 /label="ori" misc_feature 816..836 /label="Seed Sequence" CDS 7582..8385 /label="NeoR/KanR" promoter 291..296 /label="lac promoter" promoter 389..407 /label="SP6 promoter" protein_bind 329..345 /label="lac operator" promoter 315..321 /label="lac promoter" promoter 1628..1646 /label="T7 promoter" CDS 8858..9625 /label="AmpR" promoter 419..437 /label="T7 promoter" protein_bind 255..276 /label="CAP binding site" enhancer 1000..1379 /label="CMV enhancer" CDS 6247..6957 /label="mCherry" primer_bind 353..369 /label="M13 rev" CDS 8789..8857 /label="AmpR" CDS 5785..5805 /label="SV40 NLS" CDS 1666..5769 /label="Cas9" primer_bind complement(8687..8703) /label="M13 rev" misc_feature 6973..7546 /label="IRES" promoter 5854..6183 /label="SV40 promoter" promoter 297..314 /label="lac promoter" protein_bind 8711..8727 /label="lac operator" CDS 5806..5832 /label="HA" polyA_signal 8517..8638 /label="SV40 poly(A) signal" ORIGIN 1 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 61 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 121 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg 181 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 241 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt 301 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 361 agctatgaca tgattacgaa ttgcaacgat ttaggtgaca ctatagaaga gaaggaatta 421 atacgactca ctatagggag agagagagaa ttaccctcac taaagggagg agaagcatga 481 attccccagt ggaaagacgc gcaggcaaaa cgcaccacgt gacggagcgt gaccgcgcgc 541 cgagcgcgcg ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg 601 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag 661 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta 721 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa ctTGAAAGTA TTTCGATTTC 781 TTGGGTTTAT ATATCTTGTG GAAAGGACGA GGATCGCAAC TACAAGACCC GCGCCGGTTT 841 TAGAGCTAGA AATAGCAAGT TAAAATAAGG CTAGTCCGTT ATCAACTTGA AAAAGTGGCA 901 CCGAGTCGGT GCTTTTTTCT AGACACAATT GCATGAAGAA TCTGCTTAGG GTTAGGCGTT 961 TTGcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt 1021 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 1081 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 1141 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 1201 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 1261 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 1321 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg 1381 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 1441 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 1501 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 1561 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta 1621 tcgaaattaa tacgactcac tatagggaga cccaagcttg ccaccatgga caagaagtac 1681 agcatcggcc tggacatcgg taccaacagc gtgggctggg ccgtgatcac cgacgagtac 1741 aaggtgccca gcaagaagtt caaggtgctg ggcaacaccg accgccacag catcaagaag 1801 aacctgatcg gcgccctgct gttcgacagc ggcgagaccg ccgaggccac ccgcctgaag 1861 cgcaccgccc gccgccgcta cacccgccgc aagaaccgca tctgctacct gcaggagatc 1921 ttcagcaacg agatggccaa ggtggacgac agcttcttcc accgcctgga ggagagcttc 1981 ctggtggagg aggacaagaa gcacgagcgc caccccatct tcggcaacat cgtggacgag 2041 gtggcctacc acgagaagta ccccaccatc taccacctgc gcaagaagct ggtggacagc 2101 accgacaagg ccgacctgcg cctgatctac ctggccctgg cccacatgat caagttccgc 2161 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc 2221 atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc 2281 gtggacgcca aggccatcct gagcgcccgc ctgagcaaga gccgccgcct ggagaacctg 2341 atcgcccagc tgcccggcga gaagaagaac ggcctgttcg gcaacctgat cgccctgagc 2401 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc caagctgcag 2461 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag 2521 tacgccgacc tgttcctggc cgccaagaac ctgagcgacg ccatcctgct gagcgacatc 2581 ctgcgcgtga acaccgagat caccaaggcc cccctgagcg ccagcatgat caagcgctac 2641 gacgagcacc accaggacct gaccctgctg aaggccctgg tgcgccagca gctgcccgag 2701 aagtacaagg agatcttctt cgaccagagc aagaacggct acgccggcta catcgacggc 2761 ggcgccagcc aggaggagtt ctacaagttc atcaagccca tcctggagaa gatggacggc 2821 accgaggagc tgctggtgaa gctgaaccgc gaggacctgc tgcgcaagca gcgcaccttc 2881 gacaacggca gcatccccca ccagatccac ctgggcgagc tgcacgccat cctgcgccgc 2941 caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc 3001 ttccgcatcc cctactacgt gggccccctg gcccgcggca acagccgctt cgcctggatg 3061 acccgcaaga gcgaggagac catcaccccc tggaacttcg aggaggtggt ggacaagggc 3121 gccagcgccc agagcttcat cgagcgcatg accaacttcg acaagaacct gcccaacgag 3181 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc 3241 aaggtgaagt acgtgaccga gggcatgcgc aagcccgcct tcctgagcgg cgagcagaag 3301 aaggccatcg tggacctgct gttcaagacc aaccgcaagg tgaccgtgaa gcagctgaag 3361 gaggactact tcaagaagat cgagtgcttc gacagcgtgg agatcagcgg cgtggaggac 3421 cgcttcaacg ccagcctggg cacctaccac gacctgctga agatcatcaa ggacaaggac 3481 ttcctggaca acgaggagaa cgaggacatc ctggaggaca tcgtgctgac cctgaccctg 3541 ttcgaggacc gcgagatgat cgaggagcgc ctgaagacct acgcccacct gttcgacgac 3601 aaggtgatga agcagctgaa gcgccgccgc tacaccggct ggggccgcct gagccgcaag 3661 cttatcaacg gcatccgcga caagcagagc ggcaagacca tcctggactt cctgaagagc 3721 gacggcttcg ccaaccgcaa cttcatgcag ctgatccacg acgacagcct gaccttcaag 3781 gaggacatcc agaaggccca ggtgagcggc cagggcgaca gcctgcacga gcacatcgcc 3841 aacctggccg gcagccccgc catcaagaag ggcatcctgc agaccgtgaa ggtggtggac 3901 gagctggtga aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgc 3961 gagaaccaga ccacccagaa gggccagaag aacagccgcg agcgcatgaa gcgcatcgag 4021 gagggcatca aggagctggg cagccagatc ctgaaggagc accccgtgga gaacacccag 4081 ctgcagaacg agaagctgta cctgtactac ctgcagaacg gccgcgacat gtacgtggac 4141 caggagctgg acatcaaccg cctgagcgac tacgacgtgg accacatcgt gccccagagc 4201 ttcctgaagg acgacagcat cgacaacaag gtgctgaccc gcagcgacaa gaaccgcggc 4261 aagagcgaca acgtgcccag cgaggaggtg gtgaagaaga tgaagaacta ctggcgccag 4321 ctgctgaacg ccaagctgat cacccagcgc aagttcgaca acctgaccaa ggccgagcgc 4381 ggcggcctga gcgagctgga caaggccggc ttcatcaagc gccagctggt ggagacccgc 4441 cagatcacca agcacgtggc ccagatcctg gacagccgca tgaacaccaa gtacgacgag 4501 aacgacaagc tgatccgcga ggtgaaggtg atcaccctga agagcaagct ggtgagcgac 4561 ttccgcaagg acttccagtt ctacaaggtg cgcgagatca acaactacca ccacgcccac 4621 gacgcctacc tgaacgccgt ggtgggcacc gccctgatca agaagtaccc caagctggag 4681 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc gcaagatgat cgccaagagc 4741 gagcaggaga tcggcaaggc caccgccaag tacttcttct acagcaacat catgaacttc 4801 ttcaagaccg agatcaccct ggccaacggc gagatccgca agcgccccct gatcgagacc 4861 aacggcgaga ccggcgagat cgtgtgggac aagggccgcg acttcgccac cgtgcgcaag 4921 gtgctgagca tgccccaggt gaacatcgtg aagaagaccg aggtgcagac cggcggcttc 4981 agcaaggaga gcatcctgcc caagcgcaac agcgacaagc tgatcgcccg caagaaggac 5041 tgggacccca agaagtacgg cggcttcgac agccccaccg tggcctacag cgtgctggtg 5101 gtggccaagg tggagaaggg caagagcaag aagctgaaga gcgtgaagga gctgctgggc 5161 atcaccatca tggagcgcag cagcttcgag aagaacccca tcgacttcct ggaggccaag 5221 ggctacaagg aggtgaagaa ggacctgatc atcaagctgc ccaagtacag cctgttcgag 5281 ctggagaacg gccgcaagcg catgctggcc agcgccggcg agctgcagaa gggcaacgag 5341 ctggccctgc ccagcaagta cgtgaacttc ctgtacctgg ccagccacta cgagaagctg 5401 aagggcagcc ccgaggacaa cgagcagaag cagctgttcg tggagcagca caagcactac 5461 ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtgatcct ggccgacgcc 5521 aacctggaca aggtgctgag cgcctacaac aagcaccgcg acaagcccat ccgcgagcag 5581 gccgagaaca tcatccacct gttcaccctg accaacctgg gcgcccccgc cgccttcaag 5641 tacttcgaca ccaccatcga ccgcaagcgc tacaccagca ccaaggaggt gctggacgcc 5701 accctgatcc accagagcat caccggtctg tacgagaccc gcatcgacct gagccagctg 5761 ggcggcgacg gcggctccgg acctccaaag aaaaagagaa aagtataccc ctacgacgtg 5821 cccgactacg cctaataact cgagctgtgg aatgtgtgtc agttagggtg tggaaagtcc 5881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 5941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 6001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6121 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 6181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcggcgcgc cataccggtc 6241 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc 6301 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc 6361 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc 6421 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg 6481 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg 6541 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg 6601 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc 6661 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag 6721 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac 6781 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac 6841 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag 6901 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaggcg 6961 gccgcgatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga 7021 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa 7081 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc 7141 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc 7201 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg 7261 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca 7321 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tcacctcaag 7381 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct 7441 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc 7501 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc 7561 gcgataattc ctgcagccaa tatgggatcg gccattgaac aagatggatt gcacgcaggt 7621 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 7681 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 7741 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg 7801 gccacgacgg gcgttccttg cgcagcagtg ctcgacgttg tcactgaagc gggaagggac 7861 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 7921 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 7981 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 8041 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 8101 ttcgccaggc tcaaggcgcg tatgcccgac ggcgatgatc tcgtcgtgac tcatggcgat 8161 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 8221 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 8281 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 8341 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaacgcg gtgctacgag 8401 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 8461 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 8521 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 8581 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 8641 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 8701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagg 8761 taagcctgaa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 8821 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 8881 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 8941 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 9001 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 9061 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 9121 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 9181 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagggggatc atgtaactcg 9241 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 9301 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 9361 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 9421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 9481 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 9541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 9601 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 9661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 9721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 9781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 9841 aaaaccaccg ctaccagcgg tggttttttt gccggatcaa gagctaccaa ctctttttcc 9901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 9961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 10021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 10081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 10141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 10201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 10261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 10321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 10381 gaaaaacgcc agca //