LOCUS Exported 5033 bp ds-DNA circular SYN 01-JUL-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS 9169_pColEI-U6-Sp cserelheto target gRNA+BamHI-TE-mCherry SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 5033) AUTHORS Andras Talas TITLE Direct Submission JOURNAL Exported Friday, July 1, 2016 from SnapGene 3.1.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5033 /organism="recombinant plasmid" /mol_type="other DNA" promoter 7..247 /note="U6 promoter" /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 284..359 /note="gRNA scaffold" /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 743..1020 /note="chicken beta-actin promoter" CDS 1267..1977 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein" /note="mCherry" /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" polyA_signal 2106..2227 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" promoter 2490..2594 /gene="bla" /note="AmpR promoter" promoter 2596..2953 /note="SV40 promoter" /note="SV40 enhancer and early promoter" CDS 2988..3782 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin)" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" misc_feature 4014..4061 /note="HSV TK poly(A) signal" /note="herpesvirus thymidine kinase polyadenylation signal" rep_origin 4390..4978 /direction=RIGHT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt 61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga 121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac 181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg 241 aaaggacgaa acaccgggtc ttcggatcca gatctgaaga cctgttttag agctagaaat 301 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct 361 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg 421 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc 481 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg 541 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 601 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 661 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 721 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc 781 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg 841 atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg 901 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt 961 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 1021 ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc 1081 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc 1141 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt 1201 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt 1261 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc 1321 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc 1381 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc 1441 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg 1501 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg 1561 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg 1621 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc 1681 cccgtaatgc agaaaaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag 1741 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac 1801 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac 1861 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag 1921 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaggat 1981 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg 2041 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg 2101 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 2161 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 2221 tatcttaacg cggtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 2281 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 2341 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 2401 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 2461 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 2521 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 2581 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg 2641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 2701 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 2761 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 2821 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 2881 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 2941 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag 3001 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 3061 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 3121 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag 3181 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 3241 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 3301 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 3361 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 3421 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 3481 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg 3541 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 3601 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 3661 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 3721 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 3781 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 3841 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 3901 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg 3961 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 4021 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc 4081 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt 4141 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa 4201 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt 4261 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 4321 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 4381 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 4441 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 4501 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg 4561 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 4621 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 4681 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 4741 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 4801 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 4861 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 4921 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 4981 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctc //