LOCUS       sequence-450953-        6912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of GFP-FLAG tagged human Ku80 (N-terminal 
            tag). Confers resistance to G418..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2013 Jul 29."
COMMENT     Sequence Label: pEGFP-C1-FLAG-Ku80
FEATURES             Location/Qualifiers
     source          1..6912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1230..1253)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(1269..1979,1980..1982,1983..1985))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1269..1979,1980..1982,1983..1985))
                     /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(1311..1332)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1660..1679
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1919..1940
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1982..1991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2030..2233)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2059..2079)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2234..2537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2712..3300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2792..2811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3629..3676
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3701..3720
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3908..4702)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3908..4702)
                     /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(4019..4038)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4629..4648
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4737..5094)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4751..4886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4903..5094
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5051..5071
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5096..5200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5227..5682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5314..5333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5524..5545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5689..5810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5700..5719)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5754..5773
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaagccacgc cgacttgagg attagctctt ttgtcataag cgtatcgaac tatggccacc
       61 atgtctaagt catccaaagc atgaatcagg gaggaaagtg caactgcagc tgcctcatca
      121 tctcttgctg caaagacctt tagaacttga tttcccatga agaatcttct ctgaacctga
      181 gaagatttac aaaatcccaa aacagagaag cacttcccct ccgatttata tttcatttgt
      241 tcctcatcca ctttagagaa aggaactata tcacttccat agcggaaccc ttgaataata
      301 tcctctttta aaacttcagt ttcatcatca tcatttaagc aataaactgt ttctttttgt
      361 atatcttctt tttttagggt ttttgcatcc acaactgtcc aagtcttttt aactctctcc
      421 tgtagaatcg atttataggc tgcaatcctt atagacaaat tggagccaat ggtcagtcgg
      481 cagggccagt gaatggaatg cctctcaatt ttcttgaaga cgcacagttt tctcagactc
      541 tcactgaatg aataaatttc atccaaccca tcttcacctt ctaaagatat catcaccatt
      601 ttcactatct caagaccttc tttttgctgt tcggtaattc cttttagtgg aaaggaaggc
      661 ccatggccac ctaagcgaaa ggggccatct cctctgtccc cacttccatc ttccttgcca
      721 agtgagaaag gcaagaagaa ttgcagggag atgccacatt tcttcaagct atgaattata
      781 atatccagct gacttttgct gaatcggctg ctgaggtcag tgaatatttc aatatgcctc
      841 ttctcaaact tctttcctat tgtttcatgt tgaatcacat ccatgctcac gattagtgca
      901 tccaggaagt cagcctgttg agaacctggt tggattttgc tttcaatgtc ctccagcaaa
      961 tcaaaatctg gtagcatcag atgtctgtgc actgtgatgt tctgatactg atccccacca
     1021 gaaaggggat tgtcagtgcc atctgtacca aacaggacta aagcaatctc atccttgttc
     1081 tcagcaaaca cctgtcgctg tacaaacatg gttatcacct tctttgcttg ttcaaatggg
     1141 gattctatac caggaatgga gttactcatg gtaaagccca cgtccataca cagcacaaca
     1201 gctgccttat tccccgaccg cacctcgagc ttatcgtcgt catccttgta atcagatctg
     1261 agtccggact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1321 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc
     1381 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1441 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     1501 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     1561 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     1621 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     1681 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     1741 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     1801 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     1861 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     1921 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     1981 accatggtgg cgaccggtag cgctagcgga tctgacggtt cactaaacca gctctgctta
     2041 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     2101 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     2161 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     2221 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     2281 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     2341 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     2401 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     2461 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     2521 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     2581 attactatta ataactaatg catggcggta atacggttat ccacagaatc aggggataac
     2641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3421 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     3481 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     3541 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     3601 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     3661 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     3721 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     3781 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     3841 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     3901 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     3961 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     4021 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     4081 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     4141 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     4201 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4261 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4321 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     4381 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     4441 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     4501 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     4561 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     4621 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     4681 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
     4741 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     4801 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     4861 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     4921 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4981 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     5041 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
     5101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5221 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5281 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5341 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5401 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5461 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5521 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5581 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5641 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg
     5701 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5761 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5821 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     5881 aaaacctcta caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatcct
     5941 tatatcatgt ccaataaatc gtccacatca ccaccttctt caaatacagc tgctgtgtct
     6001 ccacttggtt tgtctttggg ggccagaaac tttttggctt cctcagctgt gacagaactt
     6061 ccagaggctt cctctttggt gatcagagta attccatcct ggacaacaat ttcccagaaa
     6121 tgatttaatt gtttaatctc cactttctct tgaagggctt tcaggaagtt gttaaagcgc
     6181 tgctcttctg aaaacttaat ggcttcttcc cggaaggctc ggatgcagtc tatgctcttc
     6241 ataaaatacg gtgtttcatt agtatccaaa aactgttcga tgtgatttat gagctggtta
     6301 ctcgcttcct caaagctggc cttcttctgt ttcactagaa cacggaagtt ttcagcagga
     6361 ttcacacttc caacagaggt gacactgcct tcagccagac tggagacgct gaagtgggct
     6421 cccccttgct cagtctttaa ttttttagct gtaggtccat cttcatggtt gtcttggaaa
     6481 atttcctgag cagtcacttg atcctttttc ttggcttcaa tcagaggaaa aagggtcttt
     6541 attttagaga gaggaatctg acttttcgtt gtcacctcag cgggaggatt cagcatattc
     6601 caaatatgct gctgaattgg gggtagaggc tcccggggat gtaaagctct gtgcagcaga
     6661 cactgaaata atctctgaaa tcgaggattt gggattttgg tggttggaaa caagtcttca
     6721 agggtgtctg tcttctcatc tttctttgcc aagctcatgg agtcaatcaa agcatcaaca
     6781 gcattcaact gtgcctcggt gggagcatat ttcttactgt ttttcaagga tgaaaacatg
     6841 tattgccgca agtcttccat gaaaggcagc tgcacataca ctaaacactc atagttatgc
     6901 ttgatatgag ga
//
