LOCUS       sequence-66848-e        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"
     CDS             6..29
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1204..1325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1241..1260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1295..1314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1332..1787)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1469..1490)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1681..1700
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1814..1918
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1920..2277
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1920..2111
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1943..1963)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2128..2263
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2312..3106
                     /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             2312..3106
                     /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(2366..2385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2976..2995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3294..3313)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3338..3385
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3714..4302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4203..4222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4477..4780
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4781..4984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4935..4955
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5023..5032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5029..5031,5032..5034,5035..5745)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5029..5031,5032..5034,5035..5745)
                     /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(5074..5095)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5335..5354)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5682..5703
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5761..5784
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tcgaggacta caaggacgat gacgacaagg gatccggaat ggagagaaaa ataagcagaa
       61 tccaccttgt ttctgaaccc agtataactc attttctaca agtatcttgg gagaaaacac
      121 tggaatctgg ttttgttatt acacttactg atggtcattc agcatggact gggacagttt
      181 ctgaatcaga gatttcccaa gaagctgatg acatggcaat ggaaaaaggg aaatatgttg
      241 gtgaactgag aaaagcattg ttgtcaggag caggaccagc tgatgtatac acgtttaatt
      301 tttctaaaga gtcttgttat ttcttctttg agaaaaacct gaaagatgtc tcattcagac
      361 ttggttcctt caacctagag aaagttgaaa acccagctga agtcattaga gaacttattt
      421 gttattgctt ggacaccatt gcagaaaatc aagccaaaaa tgagcacctg cagaaagaaa
      481 atgaaaggct tctgagagat tggaatgatg ttcaaggacg atttgaaaaa tgtgtgagtg
      541 ctaaggaagc tttggagact gatctttata agcggtttat tctggtgttg aatgagaaga
      601 aaacaaaaat cagaagtttg cataataaat tattaaatgc agctcaagaa cgagaaaagg
      661 acatcaaaca agaaggggaa actgcaatct gttctgaaat gactgctgac cgggaccccg
      721 tgtacgacga gagtactgat gaggaaagtg aaaaccaaac tgatctctct gggttggctt
      781 cagctgctgt aagtaaagat gattccatta tttcaagtct tgatgtcact gatattgcac
      841 caagtagaaa aaggagacag cgaatgcaaa gaaatcttgg gacagaacct aaaatggctc
      901 ctcaggagaa tcagcttcaa gaaaaggaaa agcctgattc ttcactacct gagacgtcga
      961 aaaaggagca catctcagct gaaaacatgt ctttagaaac tctgagaaac agcagcccag
     1021 aagacctctt tgatgagatt taagaattct gcagtcgacg gtaccgcggg cccgggatcc
     1081 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     1141 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1201 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1321 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1381 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1441 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1501 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1561 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1621 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1681 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1741 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     1801 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1861 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     1921 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1981 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2041 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2101 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2161 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2221 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     2281 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2341 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2401 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2461 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     2521 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2581 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2641 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2701 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2761 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2821 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2881 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2941 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3001 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3061 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3121 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3181 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3241 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     3301 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3361 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3421 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3481 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3541 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     3601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3841 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4081 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4381 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     4441 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     4501 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     4561 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     4621 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     4681 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4741 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     4801 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4861 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4921 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4981 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag
     5041 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     5101 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     5161 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     5221 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     5281 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     5341 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     5401 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     5461 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     5521 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     5581 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     5641 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     5701 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
     5761 gattacaagg atgacgacga taagc
//
