LOCUS       sequence-244398-        5342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mutant c-Jun (four N-terminal residues 
            phosphorylated by JNK are mutated into Alanines).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5342)
  AUTHORS   Aguilera C, Nakagawa K, Sancho R, Chakraborty A, Hendrich B, Behrens
            A
  TITLE     c-Jun N-terminal phosphorylation antagonises recruitment of the 
            Mbd3/NuRD repressor complex.
  JOURNAL   Nature. 2011 Jan 13;469(7329):231-5. doi: 10.1038/nature09607. Epub 
            2011 Jan 2.
  PUBMED    21196933
REFERENCE   2  (bases 1 to 5342)
  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"; doi: 
            "10.1038/nature09607"; journalName: "Nature"; date: "2011-01-13- 
            13"; volume: "469"; issue: "7329"; pages: "231-5"
COMMENT     Sequence Label: Flag-Jun4A-Myc
FEATURES             Location/Qualifiers
     source          1..5342
                     /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"
     promoter        1317..1335
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1370..1379
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1379..1402
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2438..2467
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2540..2559)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2541..2559)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2833..2954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2870..2889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2924..2943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2961..3416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3098..3119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3310..3329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3443..3547
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3549..3906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3549..3740
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3572..3592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3757..3892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3941..4735
                     /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             3941..4735
                     /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(3995..4014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4605..4624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4923..4942)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4967..5014
                     /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 attacgccaa gctcgaaatt
     1321 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ccaccatgga
     1381 ttacaaggat gacgacgata aggcccgggc ggatcccccg ggctgcagga attcgatgac
     1441 tgcaaagatg gaaacgacct tctacgacga tgccctcaac gcctcgttcc tccagtccga
     1501 gagcggtgcc tacggctaca gtaaccctaa gatcctaaaa cagagcatga ccttgaacct
     1561 ggccgacccg gtgggcagtc tgaagccgca cctccgcgcc aagaactcgg accttctcac
     1621 ggcgcccgac gtcgggctgc tcaagctggc ggcgccggag ctggagcgcc tgatcatcca
     1681 gtccagcaat gggcatatca ccactgcacc ggcccccacc cagttcttgt gccccaagaa
     1741 cgtgaccgac gagcaggagg gcttcgccga gggcttcgtg cgcgccctgg ctgaactgca
     1801 tagccagaac acgcttccca gtgtcacctc cgcggcacag ccggtcagcg gggcgggcat
     1861 ggtggctccc gcggtggcct cagtagcagg cgctggcggc ggtggtggct acagcgccag
     1921 cctgcacagt gagcctccgg tctacgccaa cctcagcaac ttcaacccgg gtgcgctgag
     1981 cagcggcggt ggggcgccct cctatggcgc ggccgggctg gcctttccct cgcagccgca
     2041 gcagcagcag cagccgcctc agccgccgca ccacttgccc caacagatcc cggtgcagca
     2101 cccgcggctg caagccctga aggaagagcc gcagaccgtg ccggagatgc cgggagagac
     2161 gccgcccctg tcccctatcg acatggagtc tcaggagcgg atcaaggcag agaggaagcg
     2221 catgaggaac cgcattgccg cctccaagtg ccggaaaagg aagctggagc ggatcgctcg
     2281 gctagaggaa aaagtgaaaa ccttgaaagc gcaaaactcc gagctggcat ccacggccaa
     2341 catgctcagg gaacaggtgg cacagcttaa gcagaaagtc atgaaccacg ttaacagtgg
     2401 gtgccaactc atgctaacgc agcagttgca aacgtttgag cagaagctga tctcggagga
     2461 ggacctatga tgaggtcgac ctcgaggggg ggcccggtac cttaattaat taaggtacca
     2521 ggtaagtgta cccaattcgc cctatagtga gtcgtattac aattcactcg atcgcccttc
     2581 ccaacagttg cgcagcctga atggcgaatg gagatccaat ttttaagtgt ataatgtgtt
     2641 aaactactga ttctaattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg
     2701 cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt
     2761 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2821 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2881 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2941 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3001 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3061 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3121 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3181 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3241 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3301 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3361 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3421 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3481 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3541 gaagaatcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3721 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3781 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3841 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3901 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3961 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4021 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4081 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4141 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4201 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4261 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4321 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4381 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gaacatcagg ggctcgcgcc
     4441 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4501 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4561 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4621 tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4681 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4741 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4801 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4861 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4921 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4981 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5041 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5101 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5161 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5221 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5281 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5341 tc
//
