LOCUS       sequence-399196-        6221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase 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 6221)
  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 6221)
  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)Luc
FEATURES             Location/Qualifiers
     source          1..6221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        416..447
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      474..483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             480..2132
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2176..2297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2187..2206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2241..2260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2491..2848
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2491..2682
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2514..2534)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2699..2834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2761..2780
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2879..3916
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3895..3903
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3940..3988
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4144..4161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4315..4903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4395..4414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5103..5894,5895..5963))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    6068..6116
                     /note="synthetic polyadenylation signal"
     misc_feature    6130..6221
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6170..6189
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcggatgggg ggtgacgtca ttcttccggg
       61 cccatattag gacttccggc ccatattagg ccgcccgcgc cggatggggg gtgacgtcat
      121 tcttccgggc ccatattagg acttccggcc catattaggc cgcccgcgcc ggatgggggg
      181 tgacgtcatt cttccgggcc catattagga cttccggccc atattaggcc gcccgcgccg
      241 gatggggggt gacgtcattc ttccgggccc atattaggac ttccggccca tattaggccg
      301 cccgcgccgg atggggggtg acgtcattct tccgggccca tattaggact tccggcccat
      361 attaggccgc ccgcgccgat atcaagatct ggcctcggcg gccaagctta gacactagag
      421 ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt aaagccacca
      481 tggaagatgc caaaaacatt aagaagggcc cagcgccatt ctacccactc gaagacggga
      541 ccgccggcga gcagctgcac aaagccatga agcgctacgc cctggtgccc ggcaccatcg
      601 cctttaccga cgcacatatc gaggtggaca ttacctacgc cgagtacttc gagatgagcg
      661 ttcggctggc agaagctatg aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt
      721 gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg
      781 ctgtggcccc agctaacgac atctacaacg agcgcgagct gctgaacagc atgggcatca
      841 gccagcccac cgtcgtattc gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa
      901 agaagctacc gatcatacaa aagatcatca tcatggatag caagaccgac taccagggct
      961 tccaaagcat gtacaccttc gtgacttccc atttgccacc cggcttcaac gagtacgact
     1021 tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct gatcatgaac agtagtggca
     1081 gtaccggatt gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc
     1141 atgcccgcga ccccatcttc ggcaaccaga tcatccccga caccgctatc ctcagcgtgg
     1201 tgccatttca ccacggcttc ggcatgttca ccacgctggg ctacttgatc tgcggctttc
     1261 gggtcgtgct catgtaccgc ttcgaggagg agctattctt gcgcagcttg caagactata
     1321 agattcaatc tgccctgctg gtgcccacac tatttagctt cttcgctaag agcactctca
     1381 tcgacaagta cgacctaagc aacttgcacg agatcgccag cggcggggcg ccgctcagca
     1441 aggaggtagg tgaggccgtg gccaaacgct tccacctacc aggcatccgc cagggctacg
     1501 gcctgacaga aacaaccagc gccattctga tcacccccga aggggacgac aagcctggcg
     1561 cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac accggtaaga
     1621 cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct
     1681 acgttaacaa ccccgaggct acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg
     1741 gcgacatcgc ctactgggac gaggacgagc acttcttcat cgtggaccgg ctgaagagcc
     1801 tgatcaaata caagggctac caggtagccc cagccgaact ggagagcatc ctgctgcaac
     1861 accccaacat cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc
     1921 ccgccgcagt cgtcgtgctg gaacacggta aaaccatgac cgagaaggag atcgtggact
     1981 atgtggccag ccaggttaca accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg
     2041 aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa gatccgcgag attctcatta
     2101 aggccaagaa gggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc
     2161 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2221 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2281 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2341 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     2401 atccgtttgc gtattgggcg ctcttccgct gatctgcgca gcaccatggc ctgaaataac
     2461 ctctgaaaga ggaacttggt tagctacctt ctgaggcgga aagaaccagc tgtggaatgt
     2521 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2581 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2641 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2701 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2761 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     2821 cttttttgga ggcctaggct tttgcaaaaa gctcgattct tctgacacta gcgccaccat
     2881 gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag
     2941 tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc cgagccttca gcttcgatgt
     3001 cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct gatggcttct acaaagaccg
     3061 ctacgtgtac cgccacttcg ccagcgctgc actacccatc cccgaagtgt tggacatcgg
     3121 cgagttcagc gagagcctga catactgcat cagtagacgc gcccaaggcg ttactctcca
     3181 agacctcccc gaaacagagc tgcctgctgt gttacagcct gtcgccgaag ctatggatgc
     3241 tattgccgcc gccgacctca gtcaaaccag cggcttcggc ccattcgggc cccaaggcat
     3301 cggccagtac acaacctggc gggatttcat ttgcgccatt gctgatcccc atgtctacca
     3361 ctggcagacc gtgatggacg acaccgtgtc cgccagcgta gctcaagccc tggacgaact
     3421 gatgctgtgg gccgaagact gtcccgaggt gcgccacctc gtccatgccg acttcggcag
     3481 caacaacgtc ctgaccgaca acggccgcat caccgccgta atcgactggt ccgaagctat
     3541 gttcggggac agtcagtacg aggtggccaa catcttcttc tggcggccct ggctggcttg
     3601 catggagcag cagactcgct acttcgagcg ccggcatccc gagctggccg gcagccctcg
     3661 tctgcgagcc tacatgctgc gcatcggcct ggatcagctc taccagagcc tcgtggacgg
     3721 caacttcgac gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc
     3781 cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga ccgacggctg
     3841 cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt acacgaccgc gcgctaagga
     3901 ggtaggtcga gtttaaactc tagaaccggt catggccgca ataaaatatc tttattttca
     3961 ttacatctgt gtgttggttt tttgtgtgtt cgaactagat gctgtcgacc gatgcccttg
     4021 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     4081 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
     4141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5041 ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa ccactgcagt
     5101 ggttaccagt gcttgatcag tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc
     5161 atagtggcct gactccccgt cgtgtagatc actacgattc gtgagggctt accatcaggc
     5221 cccagcgcag caatgatgcc gcgagagccg cgttcaccgg cccccgattt gtcagcaatg
     5281 aaccagccag cagggagggc cgagcgaaga agtggtcctg ctactttgtc cgcctccatc
     5341 cagtctatga gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag tagtttccga
     5401 agagttgtgg ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg
     5461 ttcaactctg gttcccagcg gtcaagccgg gtcacatgat cacccatatt atgaagaaat
     5521 gcagtcagct ccttagggcc tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg
     5581 ctcatggtaa tggcagcact acacaattct cttaccgtca tgccatccgt aagatgcttt
     5641 tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc
     5701 tgctcttgcc cggcgtctat acgggacaac accgcgccac atagcagtac tttgaaagtg
     5761 ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc gctattgaga
     5821 tccagttcga tatagcccac tcttgcaccc agttgatctt cagcatcttt tactttcacc
     5881 agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg aatgagtgcg
     5941 acacgaaaat gttggatgct catactcgtc ctttttcaat attattgaag catttatcag
     6001 ggttactagt acgtctctca aggataagta agtaatatta aggtacggga ggtattggac
     6061 aggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     6121 gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata
     6181 ggctgtcccc agtgcaagtg caggtgccag aacatttctc t
//
