LOCUS       sequence-193482-        6991 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Measure ERK activity by conformational change after ERK substrate 
            phosphorylation and recognition of this phosphorylated substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Demeautis C, Sipieter F, Roul J, Chapuis C, Padilla-Parra S, Riquet 
            FB, Tramier M
  TITLE     Multiplexing PKA and ERK1
  JOURNAL   Sci Rep. 2017 Jan 20;7:41026. doi: 10.1038/srep41026.
  PUBMED    28106114
REFERENCE   2  (bases 1 to 6991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2017-01-20"; volume: "7:41026. doi"; pages: " 
            10.1038/srep41026"
COMMENT     Sequence Label: EKARdual
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    34..89
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(88..107)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      177..224
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     237..942
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           325..697
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(354..942)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(951..984)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(1152..1739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1231..1250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1752..2543,2544..2612))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2375..2394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     2855..3682
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             2855..2979
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(3017..3622)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        3750..4053
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4055..4257
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4208..4228
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4460..4482
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4544..4563
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     4605..4624
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4605..4623
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4624..4648
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        4664..4773
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             4794..4811
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4821..4850
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4869..5573
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGT
                     NTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFE
                     DKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKG
                     DVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV
                     ARNSTDGMDELYK"
     CDS             join(6153..6155,6156..6158,6159..6869)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6198..6219)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6459..6478)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6806..6827
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6939..6962
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
       61 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
      121 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgtaactag
      181 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa gcatgcggag
      241 gaaattctcc ttgaagtttc cctggtgttc aaagtaaagg agtttgcacc agacgcacct
      301 ctgttcactg gtccggcgta ttaaaacacg atacattgtt attagtacat ttattaagcg
      361 ctagattctg tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta
      421 aatttataat ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta
      481 tatctgaatt taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca
      541 aacaagggtt gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc
      601 acaaaacttg ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg
      661 ttttgtaata aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca
      721 ctgtcgttag tgtacaattg actcgacgta aacacgttaa atagagcttg gacatattta
      781 acatcgggcg tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta
      841 gaagttgctt ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct
      901 aacacgtccg cgatcaaatt tgtagttgag ctttttggaa ttgcgatcgc ataacttcgt
      961 atagcataca ttatacgaag ttataagctc ggaacgctgc gctcggtcgt tcggctgcgg
     1021 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1081 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1141 ttgctggcgt ttttccatag gctccgcccc ctgacgagca tcacaaaaat cgacgctcaa
     1201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1741 aagatccttt gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2641 atttatcagg gttattgtct catgtccgcg cgtttcctgc atcttttaat caaatcccaa
     2701 gatgtgtata aacgcgccgg tatgtacagg aagaggttta tactaaactg ttacattgca
     2761 aacgtggttt cgtgtgccaa gtgtgaaaac cgatgtttaa tcaaggctct gacgcatttc
     2821 tacaaccacg actccaagtg tgtgggtgaa gtcatgcatc ttttaatcaa atcccaagat
     2881 gtgtataaac caccaaactg ccaaaaaatg aaaactgtcg acaagctctg tccgtttgct
     2941 ggcaactgca agggtctcaa tcctatttgt aattattgaa taataaaaca attataaatg
     3001 tcaaatttgt tttttattaa cgatacaaac caaacgcaac aagaacattt gtagtattat
     3061 ctataattga aaacgcgtag ttataatcgc tgaggtaata tttaaaatca ttttcaaatg
     3121 attcacagtt aatttgcgac aatataattt tattttcaca taaactagac gccttgtcgt
     3181 cttcttcttc gtattccttc tctttttcat ttttctcttc ataaaaatta acatagttat
     3241 tatcgtatcc atatatgtat ctatcgtata gagtaaattt tttgttgtca taaatatata
     3301 tgtctttttt aatggggtgt atagtaccgc tgcgcatagt ttttctgtaa tttacaacag
     3361 tgctattttc tggtagttct tcggagtgtg ttgctttaat tattaaattt atataatcaa
     3421 tgaatttggg atcgtcggtt ttgtacaata tgttgccggc atagtacgca gcttcttcta
     3481 gttcaattac accatttttt agcagcaccg gattaacata actttccaaa atgttgtacg
     3541 aaccgttaaa caaaaacagt tcacctccct tttctatact attgtctgcg agcagttgtt
     3601 tgttgttaaa aataacagcc attgtaatga gacgcacaaa ctaatatcac aaactggaaa
     3661 tgtctatcaa tatatagttg ctgatggccg gcctattaat agtaatcaat tacggggtca
     3721 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3781 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     3841 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     3901 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
     3961 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
     4021 tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca gtacaccaat
     4081 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     4141 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
     4201 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagacgtcgt
     4261 ttagtgaacc gtcagatcac tagatgcttt attgcggtag tttatcacag ttaaattgct
     4321 aacgccagtc tcgaacttaa cgtgcagaag ttggtcgtga ggcactgggc aggtaagtat
     4381 cgggcccttt gtgcgggggg agcggctcgg ggctgtccgc ggggggacgg ctgccttcgg
     4441 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     4501 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     4561 gtgctgtctc atcattttgg caaagaattg gatcggaccg aaattaatac gactcactat
     4621 aggggaattg tgagcggata acaattcccc ggagttaatc cgggaccttt aattcaaccc
     4681 aacacaatat attatagtta aataagaatt attatcaaat catttgtata ttaattaaaa
     4741 tactatactg taaattacat tttatttaca atcaaaggag atataccatg gcacaccatc
     4801 accaccacca cgggagcggg gagcagaaac taatatcaga ggaggatctt ggctcaggtt
     4861 caggatccgt gagcaagggc gaggagacca caatgggcgt aatcaagccc gacatgaaga
     4921 tcaagctgaa gatggagggc aacgtgaatg gccacgcctt cgtgatcgag ggcgagggcg
     4981 agggcaagcc ctacgacggc accaacacca tcaacctgga ggtgaaggag ggagcccccc
     5041 tgcccttctc ctacgacatt ctgaccaccg cgttcgccta cggcaacagg gccttcacca
     5101 agtaccccga cgacatcccc aactacttca agcagtcctt ccccgagggc tactcttggg
     5161 agcgcaccat gaccttcgag gacaagggca tcgtgaaggt gaagtccgac atctccatgg
     5221 aggaggactc cttcatctac gagatacacc tcaagggcga gaacttcccc cccaacggcc
     5281 ccgtgatgca gaagaagacc accggctggg acgcctccac cgagaggatg tacgtgcgcg
     5341 acggcgtgct gaagggcgac gtcaagcaca agctgctgct ggagggcggc ggccaccacc
     5401 gcgttgactt caagaccatc tacagggcca agaaggcggt gaagctgccc gactatcact
     5461 ttgtggacca ccgcatcgag atcctgaacc acgacaagga ctacaacaag gtgaccgttt
     5521 acgagagcgc cgtggcccgc aactccaccg acggcatgga cgagctgtac aagtccggaa
     5581 tggcggacga ggagaagctg ccgcccggct gggagaagcg catgagccgc agctcaggcc
     5641 gagtgtacta cttcaaccac atcactaacg ccagccagtg ggagcggccc agcggcaaca
     5701 gcagcagtgg tggcaaaaac gggcaggggg agcctgccag gaagctgcgc attctgcagt
     5761 cgactaggga gcaagggctt gtgcccgggt ctagaggcgg aggtggtgga ggaggtggag
     5821 gtggtggagg tggaggtgga ggtggtggcg gtggtggagg aggaggagct agaggcggag
     5881 gtggtggagg aggtggaggt ggtggaggtg gaggtggagg tggtggcggt ggtggaggag
     5941 gaggagccag cggcggaggt ggtggaggag gtggaggtgg tggaggtgga ggtggaggtg
     6001 gtggcggtgg tggaggagga ggaaccggtg ctgaccaact gactgaagag tggtcgacgg
     6061 tacagcggac cgcgcctgac gtaccacgta caccagtagg aaaggcaaag ctttcattcc
     6121 aattcccgcg ggcgcgggac atcgcggccg cgatggtgag caagggcgag gagctgttca
     6181 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     6241 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctcatctgca
     6301 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctga
     6361 tgtgcttcgc gcgctatcca gaccacatga agcagcacga cttcttcaag tccgccatgc
     6421 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     6481 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     6541 acttcaagga ggacggcaac atcctggggc acaagctcga gtacaactgg aacagccaca
     6601 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     6661 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     6721 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     6781 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     6841 tcactctcgg catggacgag ctgtacaagg gtaccgatat ccttcaaaaa aaacttgaag
     6901 aacttgaact tgacgaatga gatatctaat gactcgagca ccaccatcac catcaccatc
     6961 actaagtgat taacctcagg tgcaggctgc c
//
