LOCUS       sequence-396677-        4675 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 4675)
  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 4675)
  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(MapErk)DsRed
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        468..499
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      526..535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             532..1206
                     /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(662..680)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(712..732)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     930..949
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1124..1147
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1463..1584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1500..1519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1554..1573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1591..2046)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1728..1749)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1940..1959
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2073..2177
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2179..2536
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2179..2370
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2202..2222)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2387..2522
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2571..3365
                     /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             2571..3365
                     /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(2625..2644)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3235..3254
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3553..3572)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3597..3644
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3973..4561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4462..4481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagctca agcttcgaat tcggcctaac
       61 tggccggtac ctgagctcgc tagcggatgg ggggtgacgt cattcttccg ggcccatatt
      121 aggacttccg gcccatatta ggccgcccgc gccggatggg gggtgacgtc attcttccgg
      181 gcccatatta ggacttccgg cccatattag gccgcccgcg ccggatgggg ggtgacgtca
      241 ttcttccggg cccatattag gacttccggc ccatattagg ccgcccgcgc cggatggggg
      301 gtgacgtcat tcttccgggc ccatattagg acttccggcc catattaggc cgcccgcgcc
      361 ggatgggggg tgacgtcatt cttccgggcc catattagga cttccggccc atattaggcc
      421 gcccgcgccg atatcaagat ctggcctcgg cggccaagct tagacactag agggtatata
      481 atggaagctc gacttccagc ttggcaatcc ggggatccac cggtcgccac catggcctcc
      541 tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac
      601 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
      661 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
      721 cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag
      781 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
      841 gtggtgaccg tgacccagga ctcctccctg caggacggct ccttcatcta caaggtgaag
      901 ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac tatgggctgg
      961 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccacaag
     1021 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtccat ctacatggcc
     1081 aagaagcccg tgcagctgcc cggctactac tacgtggact ccaagctgga catcacctcc
     1141 cacaacgagg actacaccat cgtggagcag tacgagcgcg ccgagggccg ccaccacctg
     1201 ttcctgacta gtgatatcag ccatggcttc ccgccggcgg tggcggcgca ggatgatggc
     1261 acgctgccca tgtcttgtgc ccaggagagc gggatggacc gtcaccctgc agcctgtgct
     1321 tctgctagga tcaatgtgta ggcggccgcg actctagatc ataatcagcc ataccacatt
     1381 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1441 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1501 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1561 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     1621 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1681 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1741 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1801 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1861 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1921 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1981 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2041 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2101 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2161 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2281 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2341 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2401 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2461 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2521 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     2581 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2641 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2701 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     2761 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2821 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2881 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2941 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3001 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3061 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3121 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3181 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3241 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3301 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3361 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3421 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3481 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     3541 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     3601 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     3661 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3721 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3781 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     3841 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3901 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3961 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4021 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4081 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4141 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4201 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4261 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4321 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4381 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4441 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4501 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4561 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4621 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
