LOCUS       sequence-244361-        6927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-Daxx in mamalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  AUTHORS   Tang J, Agrawal T, Cheng Q, Qu L, Brewer MD, Chen J, Yang X
  TITLE     Phosphorylation of Daxx by ATM contributes to DNA damage-induced p53
            activation.
  JOURNAL   PLoS One. 2013;8(2):e55813. doi: 10.1371/journal.pone.0055813. Epub 
            2013 Feb 6.
  PUBMED    23405218
REFERENCE   2  (bases 1 to 6927)
  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: "PLoS One.";
            date: "2013"; volume: "8(2):e55813. doi"; pages: " 
            10.1371/journal.pone.0055813. Epub 2013 Feb 6"
COMMENT     Sequence Label: Daxx/pEGFP
FEATURES             Location/Qualifiers
     source          1..6927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /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(1361..1382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1622..1641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1969..1990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4418..4539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4455..4474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4509..4528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4546..5001)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4683..4704)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4895..4914
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5028..5132
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5134..5491
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5134..5325
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5157..5177)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5342..5477
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5526..6320
                     /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             5526..6320
                     /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(5580..5599)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6190..6209
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6508..6527)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6552..6599
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1801 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     2041 cagatccgcc accgctaaca gcatcatcgt gctggatgat gatgacgaag atgaagcagc
     2101 tgctcagcca gggccctccc acccactccc caatgcggcc tcacctgggg cagaagcccc
     2161 tagctcctct gagcctcatg gggccagagg aagcagtagt tcgggcggca agaaatgcta
     2221 caagctggag aatgagaagc tgttcgaaga gttccttgaa ctttgtaaga tgcagacagc
     2281 agaccaccct gaggtggtcc cattcctcta taaccggcag caacgtgccc actctctgtt
     2341 tttggcctcg gcggagttct gcaacatcct ctctagggtc ctgtctcggg cccggagccg
     2401 gccagccaag ctctatgtct acatcaatga gctctgcact gttctcaagg cccactcagc
     2461 caaaaagaag ctgaacttgg cccctgccgc caccacctcc aatgagccct ctgggaataa
     2521 ccctcccaca cacctctcct tggaccccac aaatgctgaa aacactgcct ctcagtctcc
     2581 aaggacccgt ggttcccggc ggcagatcca gcgtttggag cagctgctgg cgctctatgt
     2641 ggcagagatc cggcggctgc aggaaaagga gttggatctc tcagaattgg atgacccaga
     2701 ctccgcatac ctgcaggagg cacggttgaa gcgtaagctg atccgcctct ttgggcgact
     2761 atgtgagctg aaagactgct cttcactgac cggccgtgtc atagagcagc gcatccccta
     2821 ccgtggcacc cgctacccag aggttaacag gcgcattgag cggctcatca acaagccagg
     2881 gcctgatacc ttccctgact atggggatgt gcttcgggct gtagagaagg cagctgcccg
     2941 acacagcctt ggcctccccc gacagcagct ccagctcatg gctcaggatg ccttccgaga
     3001 tgtgggcatc aggttacagg agcgacgtca cctcgatctc atctacaact ttggctgcca
     3061 cctcacagat gactataggc caggcgttga ccctgcacta tcagatcctg tgttggcccg
     3121 gcgccttcgg gaaaaccgga gtttggccat gagtcggctg gatgaggtca tctccaaata
     3181 cgcaatgttg caagacaaaa gtgaggaggg cgagagaaaa aagagaagag ctcggctcca
     3241 aggcacctct tcccactctg cagacacccc cgaagcctcc ttggattctg gtgagggccc
     3301 tagtggaatg gcatcccagg ggtgcccttc tgcctccaga gctgagacag atgacgaaga
     3361 cgatgaggag agtgatgagg aagaggagga ggaggaggaa gaagaagagg aggaggccac
     3421 agattctgaa gaggaggagg atctggaaca gatgcaggag ggtcaggagg atgatgaaga
     3481 ggaggacgaa gaggaagaag cagcagcagg taaagatgga gacaagagcc ccatgtcctc
     3541 actacagatc tccaatgaaa agaacctgga acctggcaaa cagatcagca gatcttcagg
     3601 ggagcagcaa aacaaaggac gcatagtgtc accatcgtta ctgtcagaag aacccctggc
     3661 cccctccagc atagatgctg aaagcaatgg agaacagcct gaggagctga ccctggagga
     3721 agaaagccct gtgtctcagc tctttgagct agagattgaa gctttgcccc tggatacccc
     3781 ttcctctgtg gagacggaca tttcctcttc caggaagcaa tcagaggagc ccttcaccac
     3841 tgtcttagag aatggagcag gcatggtctc ttctacttcc ttcaatggag gcgtctctcc
     3901 tcacaactgg ggagattctg gtcccccctg caaaaaatct cggaaggaga agaagcaaac
     3961 aggatcaggg ccattaggaa acagctatgt ggaaaggcaa aggtcagtgc atgagaagaa
     4021 tgggaaaaag atatgtaccc tgcccagccc accttccccc ttggcttcct tggccccagt
     4081 tgctgattcc tccacgaggg tggactctcc cagccatggc ctggtgacca gctccctctg
     4141 catcccttct ccagcccggc tgtcccaaac cccccattca cagcctcctc ggcctggtac
     4201 ttgcaagaca agtgtggcca cacaatgcga tccagaagag atcatcgtgc tttcagactc
     4261 tgattaggtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     4321 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4381 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4441 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4501 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     4561 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4621 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4681 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4741 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4801 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4861 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4921 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4981 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5041 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5101 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     5161 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5221 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5281 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5341 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5401 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5461 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     5521 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5581 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5641 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5701 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5761 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5821 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5881 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5941 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6001 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     6061 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6121 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6181 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6241 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6301 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6361 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6421 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6481 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     6541 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6601 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6661 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6721 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     6781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6901 ccccgtagaa aagatcaaag gatcttc
//
