LOCUS       sequence-231166-        5785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of GFP-FLAG tagged human XRCC4 isoform 2 
            (N-terminal tag). Confers resistance to G418..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5785)
  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 5785)
  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-XRCC4
FEATURES             Location/Qualifiers
     source          1..5785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        124..427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        428..631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     582..602
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(676..678,679..681,682..1392)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(676..678,679..681,682..1392)
                     /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(721..742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(982..1001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1329..1350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1408..1431
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1438..1461
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2636..2757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2673..2692)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2727..2746
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2764..3219)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2901..2922)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3113..3132
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3246..3350
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3352..3709
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3352..3543
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3375..3395)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3560..3695
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3744..4538
                     /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             3744..4538
                     /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(3798..3817)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4408..4427
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4726..4745)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4770..4817
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5146..5734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5635..5654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
       61 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      601 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      661 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      781 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      841 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1081 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1141 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1201 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1261 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1321 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1381 gagctgtaca agtccggact cagatctgat tacaaggatg acgacgataa gctcgaggac
     1441 tacaaggacg atgacgacaa gggatccgga atggagagaa aaataagcag aatccacctt
     1501 gtttctgaac ccagtataac tcattttcta caagtatctt gggagaaaac actggaatct
     1561 ggttttgtta ttacacttac tgatggtcat tcagcatgga ctgggacagt ttctgaatca
     1621 gagatttccc aagaagctga tgacatggca atggaaaaag ggaaatatgt tggtgaactg
     1681 agaaaagcat tgttgtcagg agcaggacca gctgatgtat acacgtttaa tttttctaaa
     1741 gagtcttgtt atttcttctt tgagaaaaac ctgaaagatg tctcattcag acttggttcc
     1801 ttcaacctag agaaagttga aaacccagct gaagtcatta gagaacttat ttgttattgc
     1861 ttggacacca ttgcagaaaa tcaagccaaa aatgagcacc tgcagaaaga aaatgaaagg
     1921 cttctgagag attggaatga tgttcaagga cgatttgaaa aatgtgtgag tgctaaggaa
     1981 gctttggaga ctgatcttta taagcggttt attctggtgt tgaatgagaa gaaaacaaaa
     2041 atcagaagtt tgcataataa attattaaat gcagctcaag aacgagaaaa ggacatcaaa
     2101 caagaagggg aaactgcaat ctgttctgaa atgactgctg accgggaccc cgtgtacgac
     2161 gagagtactg atgaggaaag tgaaaaccaa actgatctct ctgggttggc ttcagctgct
     2221 gtaagtaaag atgattccat tatttcaagt cttgatgtca ctgatattgc accaagtaga
     2281 aaaaggagac agcgaatgca aagaaatctt gggacagaac ctaaaatggc tcctcaggag
     2341 aatcagcttc aagaaaagga aaagcctgat tcttcactac ctgagacgtc gaaaaaggag
     2401 cacatctcag ctgaaaacat gtctttagaa actctgagaa acagcagccc agaagacctc
     2461 tttgatgaga tttaagaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     2521 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2581 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2641 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2701 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     2761 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2821 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2881 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2941 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3001 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3061 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3121 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3181 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3241 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3301 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3361 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3421 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3481 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3541 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3601 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3661 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3721 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3781 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3841 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3901 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3961 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4021 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4081 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4141 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4201 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4261 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4321 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4381 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4441 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4501 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4561 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4621 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4681 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4741 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4801 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4861 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4921 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4981 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5281 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5401 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5461 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5521 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5581 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5641 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5701 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5761 gttcctggcc ttttgctggc ctttt
//
