LOCUS       sequence-412318-        6024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for in cell monitoring of receptor tyrosine 
            kinase-MAP ERK pathway activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6024)
  AUTHORS   Gudernova I, Foldynova-Trantirkova S, Ghannamova BE, Fafilek B, 
            Varecha M, Balek L, Hruba E, Jonatova L, Jelinkova I, Kunova 
            Bosakova M, Trantirek L, Mayer J, Krejci P
  TITLE     One reporter for in-cell activity profiling of majority of protein 
            kinase oncogenes.
  JOURNAL   Elife. 2017 Feb 15;6. pii: e21536. doi: 10.7554/eLife.21536.
  PUBMED    28199182
REFERENCE   2  (bases 1 to 6024)
  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: "Elife."; 
            date: "2017-02-15"; volume: "6. pii: e21536. doi"; pages: " 
            10.7554/eLife.21536"
COMMENT     Sequence Label: pKrox24(2xD-E)DsRed
FEATURES             Location/Qualifiers
     source          1..6024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1875..1884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1881..2555
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(2011..2029)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2061..2081)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2279..2298
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2473..2496
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2812..2933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2849..2868)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2903..2922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2940..3395)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3077..3098)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3289..3308
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3422..3526
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3528..3885
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3528..3719
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3551..3571)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3736..3871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3920..4714
                     /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             3920..4714
                     /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(3974..3993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4584..4603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4902..4921)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4946..4993
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5322..5910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5811..5830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagctca agcttggcct aactggccgg
       61 taccaaagac accgtgccat agatcgaggc ccggggtcaa ggccccgcct ctcctgggcg
      121 gcccctgccc aggcgggccc agccgctcct cccccgcact cccggttcgc tctcacggtc
      181 cctgaggtgg gcgggcgggc cctggatgac agcgatagaa ccccggcccg actcgccctc
      241 gcccccgctc tgggtctggg cttccccagc ctagttcacg cctaggagcc gcctgagcag
      301 ccgcgcgccc agcgccacac gccacgagcc ctccccgcct gggcgtcccc ggatcccgcg
      361 agcgctcggg ctcccggctt ggaaccaggg aggagggagg gagcgaggga gcaaccagct
      421 gcgacccgga aatgccatat aaggagcagg aaggatcccc cgccggaaag acaccgtgcc
      481 atagatcgag gcccggggtc aaggccccgc ctctcctggg cggcccctgc ccaggcgggc
      541 ccagccgctc ctcccccgca ctcccggttc gctctcacgg tccctgaggt gggcgggcgg
      601 gccctggatg acagcgatag aaccccggcc cgactcgccc tcgcccccgc tctgggtctg
      661 ggcttcccca gcctagttca cgcctaggag ccgcctgagc agccgcgcgc ccagcgccac
      721 acgccacgag ccctccccgc ctgggcgtcc ccggatcccg cgagcgctcg ggctcccggc
      781 ttggaaccag ggaggaggga gggagcgagg gagcaaccag ctgcgacccg gaaatgccat
      841 ataaggagca ggaaggatcc cccgccggga gctcggtacc aaagacaccg tgccatagat
      901 cgaggcccgg ggtcaaggcc ccgcctctcc tgggcggccc ctgcccaggc gggcccagcc
      961 gctcctcccc cgcactcccg gttcgctctc acggtccctg aggtgggcgg gcgggccctg
     1021 ggtgacagcg atagaacccc ggcccgactc gccctcgccc ccgctctggg tctgggcttc
     1081 cccagcctag ttcacgccta ggagccgcct gagcagccgc gcgcccagcg ccacacgcca
     1141 cgagccctcc ccgcctgggc gtccccggat cccgcgagcg ctcgggctcc cggcttggaa
     1201 ccagggagga gggagggagc gagggagcaa ccagctgcga cccggaaatg ccatataagg
     1261 agcaggaagg atcccccgcc ggaacaaccc ttatttgggc agcaccttat ttggagtggc
     1321 ccgatatggc ccggccgctt ccggctctgg gaggagggaa gaaggcggag ggaggggcaa
     1381 cgcgggaact ccggagctgc gcgggtcccg gaggccccgg cggcggctag agctctaggc
     1441 ttccccgaag cctgggcgcc tgggatgcgg gcgcgggcgc gggccctagg gtgcaggatg
     1501 gaggtgccgg gcgctgtcgg atggggggct tcacgtcact ccgggtcctc ccggccggtc
     1561 ctgccatatt agggcttcct gcttcccata tatggccatg tacgtcacga cggaggcgga
     1621 cccgtgccgt tccagaccct tcaaatagag gcggatccgg ggagtcgcga gagatcccag
     1681 cgcgcagaac ttggggagcc gccgccgcca tccgccgccg cagccagctt ccgccgccgc
     1741 aggaccggcc cctgccccag cctccgcagc cgcggcgcgt ccacgcccgc ccgcgcccag
     1801 ggcgagtcgg ggtcgccgcc tgcacgcttc tcagtgttcc gtcgacggta ccgcgggccc
     1861 gggatccacc ggtcgccacc atggcctcct ccgaggacgt catcaaggag ttcatgcgct
     1921 tcaaggtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     1981 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     2041 tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga
     2101 agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg
     2161 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     2221 aggacggctc cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc
     2281 ccgtaatgca gaagaagact atgggctggg aggcctccac cgagcgcctg tacccccgcg
     2341 acggcgtgct gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc
     2401 tggtggagtt caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact
     2461 acgtggactc caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt
     2521 acgagcgcgc cgagggccgc caccacctgt tcctgactag tgatatcagc catggcttcc
     2581 cgccggcggt ggcggcgcag gatgatggca cgctgcccat gtcttgtgcc caggagagcg
     2641 ggatggaccg tcaccctgca gcctgtgctt ctgctaggat caatgtgtag gcggccgcga
     2701 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2761 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2821 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2881 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2941 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3001 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3061 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3121 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3181 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3241 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3301 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3361 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3421 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3541 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3601 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3721 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3781 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3841 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3901 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3961 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4021 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4081 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4141 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4201 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4261 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4321 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4381 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4441 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4501 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4561 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4621 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4681 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4741 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4801 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4861 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4921 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4981 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5041 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5101 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5161 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5221 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5281 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5341 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5401 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5461 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5521 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5581 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5641 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5701 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5761 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5821 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5881 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5941 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6001 gataaccgta ttaccgccat gcat
//
