LOCUS Construct_7 7956 bp DNA circular UNA 20-APR-2016 DEFINITION CMVp-Cas9-3xNLS-HSVpA. ACCESSION urn.local...1461189080499.38 VERSION urn.local...1461189080499.38 KEYWORDS . SOURCE Human ORGANISM Human . FEATURES Location/Qualifiers gene complement(451..1311) /created_by="User" /label="AmpR" rep_origin 1443..1898 /created_by="User" /label="phage f1 origin of replication" promoter 2027..2615 /Original_Bases /modified_by="User" /label="pmCMV" sig_peptide 2636..2641 /created_by="User" /label="Kozak" CDS 2642..6745 /note="hSpCas9" /modified_by="User" /label="hSpCas9" primer_bind 3205..3232 /Mismatches=0 /created_by="primer3" /annotation_group="1413859499580818000240e988e-0722-4220-b 424-f067d74405ed" /%GC="57.1" /Tm="68.8" /Hairpin_Tm="39.5" /Self_Dimer_Tm="3.5" /Sequence="GGTGCAGACCTACAACCAGCTGTTCGAG" /label="JP1663" primer_bind 3871..3901 /Mismatches=0 /created_by="primer3" /annotation_group="1413859499810770000c4f7a1e7-6365-4417-8 7cb-b77007ef64eb" /%GC="67.7" /Tm="75.5" /Hairpin_Tm="53.8" /Self_Dimer_Tm="21.3" /Sequence="CCCCCACCAGATCCACCTGGGAGAGCTGCAC" /label="JP1664" primer_bind 4477..4510 /Mismatches=0 /created_by="primer3" /annotation_group="1413859500041800000668f4874-ab7e-4f5c-a 9e0-0c91871aa3ae" /%GC="52.9" /Tm="70.5" /Hairpin_Tm="None" /Self_Dimer_Tm="14.4" /Sequence="CGAGGACATTCTGGAAGATATCGTGCTGACCCTG" /label="JP1665" primer_bind 5107..5140 /Mismatches=0 /modified_by="User" /created_by="primer3" /annotation_group="14138595002407990008b8a2999-9296-4397-a 525-0994bba8d18f" /%GC="58.8" /Tm="73.7" /Hairpin_Tm="78.4" /Self_Dimer_Tm="13.9" /Sequence="GTACGTGGACCAGGAACTGGACATCAACCGGCTG" /label="JP1666" primer_bind 5784..5815 /Mismatches=0 /modified_by="User" /created_by="primer3" /annotation_group="141385950047921100052c3b3a9-66ea-44a0-b 66e-1d0326184345" /%GC="65.6" /Tm="75.4" /Hairpin_Tm="53.7" /Self_Dimer_Tm="19.1" /Sequence="CCGAGATTACCCTGGCCAACGGCGAGATCCGG" /label="JP1667" residue 6005..6007 /created_by="User" /label="Arg1122" primer_bind 6484..6517 /Mismatches=0 /modified_by="User" /created_by="primer3" /annotation_group="14138595006972170005bf14214-cb3c-4c08-8 bc9-7cdcb439bedb" /%GC="58.8" /Tm="74.1" /Hairpin_Tm="48.4" /Self_Dimer_Tm="15.0" /Sequence="CCTGGCCGACGCTAATCTGGACAAAGTGCTGTCC" /label="JP1668" misc_signal 6746..6793 /created_by="User" /modified_by="User" /label="NLS" misc_signal 6818..6838 /created_by="User" /label="NLS" misc_signal 6845..6865 /created_by="User" /label="NLS" polyA_signal 6878..7327 /Original_Bases /modified_by="User" /label="HSV polyA" ORIGIN 1 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 61 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 121 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 421 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 481 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 541 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 601 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 661 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 721 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 781 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 841 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 901 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 961 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 1021 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 1081 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 1141 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 1201 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 1261 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 1321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 1381 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 1441 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 1501 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 1561 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 1621 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 1681 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 1741 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 1801 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 1861 taacgcgaat tttaacaaaa tattaacgtt tacaatttcc cattcgccat tcaggctgcg 1921 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc 1981 atgataagta agtaatatta aggtacggga ggtacttgga ggtacctagt tattaatagt 2041 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 2101 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 2161 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 2221 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 2281 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 2341 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt 2401 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 2461 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 2521 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 2581 atataagcag agctggttta tgaaccgtca gatccgagct catcaccggt gcgctgccac 2641 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt 2701 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg 2761 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga 2821 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg 2881 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag 2941 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg 3001 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa 3061 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca 3121 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga 3181 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc 3241 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag 3301 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa 3361 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga 3421 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc 3481 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat 3541 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc 3601 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg 3661 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc 3721 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct 3781 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg 3841 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca 3901 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat 3961 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag 4021 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga 4081 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa 4141 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt 4201 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct 4261 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac 4321 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat 4381 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat 4441 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt 4501 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc 4561 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg 4621 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct 4681 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga 4741 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct 4801 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac 4861 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt 4921 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag 4981 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc 5041 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg 5101 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca 5161 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag 5221 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa 5281 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct 5341 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca 5401 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa 5461 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc 5521 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa 5581 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa 5641 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa 5701 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag 5761 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg 5821 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt 5881 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt 5941 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat 6001 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc 6061 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt 6121 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga 6181 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa 6241 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact 6301 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag 6361 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga 6421 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt 6481 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa 6541 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc 6601 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa 6661 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat 6721 cgacctgtct cagctgggag gcgacaagcg tcctgctgct actaagaaag ctggtcaagc 6781 taagaaaaag aaagctagcg gcagcggcgc cggatcccca aagaagaaaa ggaaggttga 6841 agaccccaag aaaaagagga aggtgatacc cgggtaagcg ggactctggg gttcgaaatg 6901 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 6961 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 7021 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag 7081 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt 7141 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc 7201 ccaccgagac gccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc 7261 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata 7321 gcctcagctc gagtatgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc 7381 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa 7441 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7561 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 7861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7921 gcccgaccgc tgcgccttat ccggtaacta tcgtct //