LOCUS Exported 6543 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid for cloning and epression of GFP-FLAG tagged human Ku70 (N-terminal tag). Confers resistance to G418.. ACCESSION . VERSION . KEYWORDS pEGFP-C1-FLAG-Ku70 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6543) AUTHORS Britton S, Coates J, Jackson SP TITLE A new method for high-resolution imaging of Ku foci to decipher mechanisms of DNA double-strand break repair. JOURNAL J Cell Biol. 2013 Jul 29. PUBMED 23897892 REFERENCE 2 (bases 1 to 6543) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6543 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 1962..2083 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1999..2018) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2053..2072 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2090..2545) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2227..2248) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2439..2458 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2572..2676 /gene="bla" /label=AmpR promoter promoter 2678..3035 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2701..2721) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2886..3021 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2948..2967 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3070..3864 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3124..3143) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3734..3753 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4052..4071) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4096..4143 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4472..5060 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4961..4980 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 5235..5538 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5539..5742 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5693..5713 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5781..5790 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5787..6503 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 5787..6503 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5832..5853) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6093..6112) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6440..6461 /label=EGFP-C /note="EGFP, forward primer" CDS 6519..6542 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" ORIGIN 1 tcgagtcagg gtgggagtca tattacaaaa ccgagggcga tgaagaagca gaggaagaac 61 aagaagagaa ccttgaagca agtggagact ataaatattc aggaagagat agtttgattt 121 ttttggttga tgcctccaag gctatgtttg aatctcagag tgaagatgag ttgacacctt 181 ttgacatgag catccagtgt atccaaagtg tgtacatcag taagatcata agcagtgatc 241 gagatctctt ggctgtggtg ttctatggta ccgagaaaga caaaaattca gtgaatttta 301 aaaatattta cgtcttacag gagctggata atccaggtgc aaaacgaatt ctagagcttg 361 accagtttaa ggggcagcag ggacaaaaac gtttccaaga catgatgggc cacggatctg 421 actactcact cagtgaagtg ctgtgggtct gtgccaacct ctttagtgat gtccaattca 481 agatgagtca taagaggatc atgctgttca ccaatgaaga caacccccat ggcaatgaca 541 gtgccaaagc cagccgggcc aggaccaaag ccggtgatct ccgagataca ggcatcttcc 601 ttgacttgat gcacctgaag aaacctgggg gctttgacat atccttgttc tacagagata 661 tcatcagcat agcagaggat gaggacctca gggttcactt tgaggaatcc agcaagctag 721 aagacctgtt gcggaaggtt cgcgccaagg agacaaggaa gcgagcactc agcaggttaa 781 agctgaagct caacaaagat atagtgatct ctgtgggcat ttataatctg gtccagaagg 841 ctctcaagcc tcctccaata aagctctatc gggaaacaaa tgaaccagtg aaaaccaaga 901 cccggacctt taatacaagt acaggcggtt tgcttctgcc tagcgatacc aagaggtctc 961 agatctatgg gagtcgtcag attatactgg agaaagagga aacagaagag ctaaaacggt 1021 ttgatgatcc aggtttgatg ctcatgggtt tcaagccgtt ggtactgctg aagaaacacc 1081 attacctgag gccctccctg ttcgtgtacc cagaggagtc gctggtgatt gggagctcaa 1141 ccctgttcag tgctctgctc atcaagtgtc tggagaagga ggttgcagca ttgtgcagat 1201 acacaccccg caggaacatc cctccttatt ttgtggcttt ggtgccacag gaagaagagt 1261 tggatgacca gaaaattcag gtgactcctc caggcttcca gctggtcttt ttaccctttg 1321 ctgatgataa aaggaagatg ccctttactg aaaaaatcat ggcaactcca gagcaggtgg 1381 gcaagatgaa ggctatcgtt gagaagcttc gcttcacata cagaagtgac agctttgaga 1441 accccgtgct gcagcagcac ttcaggaacc tggaggcctt ggccttggat ttgatggagc 1501 cggaacaagc agtggacctg acattgccca aggttgaagc aatgaataaa agactgggct 1561 ccttggtgga tgagtttaag gagcttgttt acccaccaga ttacaatcct gaagggaaag 1621 ttaccaagag aaaacacgat aatgaaggtt ctggaagcaa aaggcccaag gtggagtatt 1681 cagaagagga gctgaagacc cacatcagca agggtacgct gggcaagttc actgtgccca 1741 tgctgaaaga ggcctgccgg gcttacgggc tgaagagtgg gctgaagaag caggagctgc 1801 tggaagccct caccaagcac ttccaggact aaggatccac cggatctaga taactgatca 1861 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 1921 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 1981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 2041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc 2101 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 2161 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 2221 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 2281 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 2341 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 2401 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 2461 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 2521 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 2581 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 2641 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt 2701 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2761 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 2821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 2881 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 2941 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 3001 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga 3061 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 3121 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 3181 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 3241 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 3301 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 3361 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 3421 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 3481 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 3541 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc 3601 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 3661 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 3721 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 3781 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 3841 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 3901 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 3961 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 4021 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 4081 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg 4141 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc 4201 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg 4261 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact 4321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 4381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 4441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 4501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 4561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc 4621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 4681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 4741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 4801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 4861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 4921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 4981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 5041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 5101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 5161 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 5221 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 5281 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 5341 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 5401 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 5461 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 5521 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 5581 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 5641 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 5701 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag 5761 atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg 5821 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 5881 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 5941 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 6001 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 6061 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 6121 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 6181 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 6241 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 6301 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 6361 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 6421 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 6481 tcggcatgga cgagctgtac aagtccggac tcagatctga ttacaaggat gacgacgata 6541 agc //