LOCUS       Exported                6912 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  Plasmid for expression of GFP-FLAG tagged human Ku80 (N-terminal 
            tag). Confers resistance to G418..
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-C1-FLAG-Ku80
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6912)
  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 6912)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 20, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6912
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(24..43)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     270..290
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(315..419)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      446..901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(533..552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     743..764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    908..1029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(919..938)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     973..992
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3361..3384)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(3400..4116)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(3400..4116)
                     /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(3442..3463)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3791..3810
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4050..4071
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4113..4122
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(4161..4364)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4190..4210)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4365..4668
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4843..5431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4923..4942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5760..5807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5832..5851
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6039..6833)
                     /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(6150..6169)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6760..6779
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6868..313)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6882..105
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
       61 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
      121 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
      181 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
      241 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
      301 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      421 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
      481 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      541 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      601 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      661 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      721 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      781 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      841 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
      901 tacgcgttaa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
      961 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1021 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1081 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtta
     1141 tctagatccg gtggatcctt atatcatgtc caataaatcg tccacatcac caccttcttc
     1201 aaatacagct gctgtgtctc cacttggttt gtctttgggg gccagaaact ttttggcttc
     1261 ctcagctgtg acagaacttc cagaggcttc ctctttggtg atcagagtaa ttccatcctg
     1321 gacaacaatt tcccagaaat gatttaattg tttaatctcc actttctctt gaagggcttt
     1381 caggaagttg ttaaagcgct gctcttctga aaacttaatg gcttcttccc ggaaggctcg
     1441 gatgcagtct atgctcttca taaaatacgg tgtttcatta gtatccaaaa actgttcgat
     1501 gtgatttatg agctggttac tcgcttcctc aaagctggcc ttcttctgtt tcactagaac
     1561 acggaagttt tcagcaggat tcacacttcc aacagaggtg acactgcctt cagccagact
     1621 ggagacgctg aagtgggctc ccccttgctc agtctttaat tttttagctg taggtccatc
     1681 ttcatggttg tcttggaaaa tttcctgagc agtcacttga tcctttttct tggcttcaat
     1741 cagaggaaaa agggtcttta ttttagagag aggaatctga cttttcgttg tcacctcagc
     1801 gggaggattc agcatattcc aaatatgctg ctgaattggg ggtagaggct cccggggatg
     1861 taaagctctg tgcagcagac actgaaataa tctctgaaat cgaggatttg ggattttggt
     1921 ggttggaaac aagtcttcaa gggtgtctgt cttctcatct ttctttgcca agctcatgga
     1981 gtcaatcaaa gcatcaacag cattcaactg tgcctcggtg ggagcatatt tcttactgtt
     2041 tttcaaggat gaaaacatgt attgccgcaa gtcttccatg aaaggcagct gcacatacac
     2101 taaacactca tagttatgct tgatatgagg aaaagccacg ccgacttgag gattagctct
     2161 tttgtcataa gcgtatcgaa ctatggccac catgtctaag tcatccaaag catgaatcag
     2221 ggaggaaagt gcaactgcag ctgcctcatc atctcttgct gcaaagacct ttagaacttg
     2281 atttcccatg aagaatcttc tctgaacctg agaagattta caaaatccca aaacagagaa
     2341 gcacttcccc tccgatttat atttcatttg ttcctcatcc actttagaga aaggaactat
     2401 atcacttcca tagcggaacc cttgaataat atcctctttt aaaacttcag tttcatcatc
     2461 atcatttaag caataaactg tttctttttg tatatcttct ttttttaggg tttttgcatc
     2521 cacaactgtc caagtctttt taactctctc ctgtagaatc gatttatagg ctgcaatcct
     2581 tatagacaaa ttggagccaa tggtcagtcg gcagggccag tgaatggaat gcctctcaat
     2641 tttcttgaag acgcacagtt ttctcagact ctcactgaat gaataaattt catccaaccc
     2701 atcttcacct tctaaagata tcatcaccat tttcactatc tcaagacctt ctttttgctg
     2761 ttcggtaatt ccttttagtg gaaaggaagg cccatggcca cctaagcgaa aggggccatc
     2821 tcctctgtcc ccacttccat cttccttgcc aagtgagaaa ggcaagaaga attgcaggga
     2881 gatgccacat ttcttcaagc tatgaattat aatatccagc tgacttttgc tgaatcggct
     2941 gctgaggtca gtgaatattt caatatgcct cttctcaaac ttctttccta ttgtttcatg
     3001 ttgaatcaca tccatgctca cgattagtgc atccaggaag tcagcctgtt gagaacctgg
     3061 ttggattttg ctttcaatgt cctccagcaa atcaaaatct ggtagcatca gatgtctgtg
     3121 cactgtgatg ttctgatact gatccccacc agaaagggga ttgtcagtgc catctgtacc
     3181 aaacaggact aaagcaatct catccttgtt ctcagcaaac acctgtcgct gtacaaacat
     3241 ggttatcacc ttctttgctt gttcaaatgg ggattctata ccaggaatgg agttactcat
     3301 ggtaaagccc acgtccatac acagcacaac agctgcctta ttccccgacc gcacctcgag
     3361 cttatcgtcg tcatccttgt aatcagatct gagtccggac ttgtacagct cgtccatgcc
     3421 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     3481 ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
     3541 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     3601 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     3661 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     3721 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     3781 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     3841 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     3901 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     3961 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     4021 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     4081 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta gcgctagcgg
     4141 atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg
     4201 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     4261 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     4321 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     4381 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     4441 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     4501 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     4561 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     4621 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     4681 tatgggctat gaactaatga ccccgtaatt gattactatt aataactaat gcatggcggt
     4741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5581 tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc
     5641 ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca
     5701 atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg
     5761 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     5821 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa
     5881 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     5941 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     6001 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     6061 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     6121 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     6181 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     6241 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     6301 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     6361 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     6421 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     6481 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     6541 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     6601 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     6661 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     6721 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     6781 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     6841 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
     6901 tcactacttc tg
//
