LOCUS       Exported                6912 bp ds-DNA     circular SYN 12-MAY-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 May 12, 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"
     polyA_signal    2331..2452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2368..2387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2422..2441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2459..2914)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2596..2617)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2808..2827
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2941..3045
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3047..3404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3070..3090)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3255..3390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3317..3336
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3439..4233
                     /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(3493..3512)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4103..4122
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4421..4440)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4465..4512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4841..5429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5330..5349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5604..5907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5908..6111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6062..6082
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6150..6159
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6156..6872
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             6156..6872
                     /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(6201..6222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6462..6481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6809..6830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6888..6911
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tcgaggtgcg gtcggggaat aaggcagctg ttgtgctgtg tatggacgtg ggctttacca
       61 tgagtaactc cattcctggt atagaatccc catttgaaca agcaaagaag gtgataacca
      121 tgtttgtaca gcgacaggtg tttgctgaga acaaggatga gattgcttta gtcctgtttg
      181 gtacagatgg cactgacaat cccctttctg gtggggatca gtatcagaac atcacagtgc
      241 acagacatct gatgctacca gattttgatt tgctggagga cattgaaagc aaaatccaac
      301 caggttctca acaggctgac ttcctggatg cactaatcgt gagcatggat gtgattcaac
      361 atgaaacaat aggaaagaag tttgagaaga ggcatattga aatattcact gacctcagca
      421 gccgattcag caaaagtcag ctggatatta taattcatag cttgaagaaa tgtgacatct
      481 ccctgcaatt cttcttgcct ttctcacttg gcaaggaaga tggaagtggg gacagaggag
      541 atggcccctt tcgcttaggt ggccatgggc cttcctttcc actaaaagga attaccgaac
      601 agcaaaaaga aggtcttgag atagtgaaaa tggtgatgat atctttagaa ggtgaagatg
      661 ggttggatga aatttattca ttcagtgaga gtctgagaaa actgtgcgtc ttcaagaaaa
      721 ttgagaggca ttccattcac tggccctgcc gactgaccat tggctccaat ttgtctataa
      781 ggattgcagc ctataaatcg attctacagg agagagttaa aaagacttgg acagttgtgg
      841 atgcaaaaac cctaaaaaaa gaagatatac aaaaagaaac agtttattgc ttaaatgatg
      901 atgatgaaac tgaagtttta aaagaggata ttattcaagg gttccgctat ggaagtgata
      961 tagttccttt ctctaaagtg gatgaggaac aaatgaaata taaatcggag gggaagtgct
     1021 tctctgtttt gggattttgt aaatcttctc aggttcagag aagattcttc atgggaaatc
     1081 aagttctaaa ggtctttgca gcaagagatg atgaggcagc tgcagttgca ctttcctccc
     1141 tgattcatgc tttggatgac ttagacatgg tggccatagt tcgatacgct tatgacaaaa
     1201 gagctaatcc tcaagtcggc gtggcttttc ctcatatcaa gcataactat gagtgtttag
     1261 tgtatgtgca gctgcctttc atggaagact tgcggcaata catgttttca tccttgaaaa
     1321 acagtaagaa atatgctccc accgaggcac agttgaatgc tgttgatgct ttgattgact
     1381 ccatgagctt ggcaaagaaa gatgagaaga cagacaccct tgaagacttg tttccaacca
     1441 ccaaaatccc aaatcctcga tttcagagat tatttcagtg tctgctgcac agagctttac
     1501 atccccggga gcctctaccc ccaattcagc agcatatttg gaatatgctg aatcctcccg
     1561 ctgaggtgac aacgaaaagt cagattcctc tctctaaaat aaagaccctt tttcctctga
     1621 ttgaagccaa gaaaaaggat caagtgactg ctcaggaaat tttccaagac aaccatgaag
     1681 atggacctac agctaaaaaa ttaaagactg agcaaggggg agcccacttc agcgtctcca
     1741 gtctggctga aggcagtgtc acctctgttg gaagtgtgaa tcctgctgaa aacttccgtg
     1801 ttctagtgaa acagaagaag gccagctttg aggaagcgag taaccagctc ataaatcaca
     1861 tcgaacagtt tttggatact aatgaaacac cgtattttat gaagagcata gactgcatcc
     1921 gagccttccg ggaagaagcc attaagtttt cagaagagca gcgctttaac aacttcctga
     1981 aagcccttca agagaaagtg gagattaaac aattaaatca tttctgggaa attgttgtcc
     2041 aggatggaat tactctgatc accaaagagg aagcctctgg aagttctgtc acagctgagg
     2101 aagccaaaaa gtttctggcc cccaaagaca aaccaagtgg agacacagca gctgtatttg
     2161 aagaaggtgg tgatgtggac gatttattgg acatgatata aggatccacc ggatctagat
     2221 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2281 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     2461 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2521 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2581 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2641 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2701 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2761 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2821 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2881 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2941 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3001 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3061 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3601 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3961 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4381 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4441 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4501 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4561 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4621 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4681 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4981 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5521 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     5581 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5641 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5701 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     5761 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     5821 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5881 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     5941 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6001 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6061 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     6121 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     6181 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     6241 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     6301 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     6361 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     6421 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     6481 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     6541 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     6601 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     6661 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     6721 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     6781 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     6841 ggatcactct cggcatggac gagctgtaca agtccggact cagatctgat tacaaggatg
     6901 acgacgataa gc
//
