LOCUS       sequence-415367-        7359 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 7359)
  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 7359)
  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_inD)Luc
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1854..1863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1860..3512
                     /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    3556..3677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3567..3586)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3621..3640
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3871..4228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3871..4062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3894..3914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4079..4214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4141..4160
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4259..5053
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5078..5126
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5282..5299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5453..6041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5533..5552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6241..7032,7033..7101))
                     /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    7206..7254
                     /note="synthetic polyadenylation signal"
     misc_feature    7268..7359
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7308..7327
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca aagacaccgt gccatagatc gaggcccggg gtcaaggccc
       61 cgcctctcct gggcggcccc tgcccaggcg ggcccagccg ctcctccccc gcactcccgg
      121 ttcgctctca cggtccctga ggtgggcggg cgggccctgg atgacagcga tagaaccccg
      181 gcccgactcg ccctcgcccc cgctctgggt ctgggcttcc ccagcctagt tcacgcctag
      241 gagccgcctg agcagccgcg cgcccagcgc cacacgccac gagccctccc cgcctgggcg
      301 tccccggatc ccgcgagcgc tcgggctccc ggcttggaac cagggaggag ggagggagcg
      361 agggagcaac cagctgcgac ccggaaatgc catataagga gcaggaagga tcccccgccg
      421 gaaagacacc gtgccataga tcgaggcccg gggtcaaggc cccgcctctc ctgggcggcc
      481 cctgcccagg cgggcccagc cgctcctccc ccgcactccc ggttcgctct cacggtccct
      541 gaggtgggcg ggcgggccct ggatgacagc gatagaaccc cggcccgact cgccctcgcc
      601 cccgctctgg gtctgggctt ccccagccta gttcacgcct aggagccgcc tgagcagccg
      661 cgcgcccagc gccacacgcc acgagccctc cccgcctggg cgtccccgga tcccgcgagc
      721 gctcgggctc ccggcttgga accagggagg agggagggag cgagggagca accagctgcg
      781 acccggaaat gccatataag gagcaggaag gatcccccgc cgggagctcg gtaccaaaga
      841 caccgtgcca tagatcgagg cccggggtca aggccccgcc tctcctgggc ggcccctgcc
      901 caggcgggcc cagccgctcc tcccccgcac tcccggttcg ctctcacggt ccctgaggtg
      961 ggcgggcggg ccctgggtga cagcgataga accccggccc gactcgccct cgcccccgct
     1021 ctgggtctgg gcttccccag cctagttcac gcctaggagc cgcctgagca gccgcgcgcc
     1081 cagcgccaca cgccacgagc cctccccgcc tgggcgtccc cggatcccgc gagcgctcgg
     1141 gctcccggct tggaaccagg gaggagggag ggagcgaggg agcaaccagc tgcgacccgg
     1201 aaatgccata taaggagcag gaaggatccc ccgccggaac aacccttatt tgggcagcac
     1261 cttatttgga gtggcccgat atggcccggc cgcttccggc tctgggagga gggaagaagg
     1321 cggagggagg ggcaacgcgg gaactccgga gctgcgcggg tcccggaggc cccggcggcg
     1381 gctagagctc taggcttccc cgaagcctgg gcgcctggga tgcgggcgcg ggcgcgggcc
     1441 ctagggtgca ggatggaggt gccgggcgct gtcggatggg gggcttcacg tcactccggg
     1501 tcctcccggc cggtcctgcc atattagggc ttcctgcttc ccatatatgg ccatgtacgt
     1561 cacgacggag gcggacccgt gccgttccag acccttcaaa tagaggcgga tccggggagt
     1621 cgcgagagat cccagcgcgc agaacttggg gagccgccgc cgccatccgc cgccgcagcc
     1681 agcttccgcc gccgcaggac cggcccctgc cccagcctcc gcagccgcgg cgcgtccacg
     1741 cccgcccgcg cccagggcga gtcggggtcg ccgcctgcac gcttctcagt gttccgatat
     1801 catcaagatc tggcctcggc ggccaagctt ggcaatccgg tactgttggt aaagccacca
     1861 tggaagatgc caaaaacatt aagaagggcc cagcgccatt ctacccactc gaagacggga
     1921 ccgccggcga gcagctgcac aaagccatga agcgctacgc cctggtgccc ggcaccatcg
     1981 cctttaccga cgcacatatc gaggtggaca ttacctacgc cgagtacttc gagatgagcg
     2041 ttcggctggc agaagctatg aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt
     2101 gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg
     2161 ctgtggcccc agctaacgac atctacaacg agcgcgagct gctgaacagc atgggcatca
     2221 gccagcccac cgtcgtattc gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa
     2281 agaagctacc gatcatacaa aagatcatca tcatggatag caagaccgac taccagggct
     2341 tccaaagcat gtacaccttc gtgacttccc atttgccacc cggcttcaac gagtacgact
     2401 tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct gatcatgaac agtagtggca
     2461 gtaccggatt gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc
     2521 atgcccgcga ccccatcttc ggcaaccaga tcatccccga caccgctatc ctcagcgtgg
     2581 tgccatttca ccacggcttc ggcatgttca ccacgctggg ctacttgatc tgcggctttc
     2641 gggtcgtgct catgtaccgc ttcgaggagg agctattctt gcgcagcttg caagactata
     2701 agattcaatc tgccctgctg gtgcccacac tatttagctt cttcgctaag agcactctca
     2761 tcgacaagta cgacctaagc aacttgcacg agatcgccag cggcggggcg ccgctcagca
     2821 aggaggtagg tgaggccgtg gccaaacgct tccacctacc aggcatccgc cagggctacg
     2881 gcctgacaga aacaaccagc gccattctga tcacccccga aggggacgac aagcctggcg
     2941 cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac accggtaaga
     3001 cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct
     3061 acgttaacaa ccccgaggct acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg
     3121 gcgacatcgc ctactgggac gaggacgagc acttcttcat cgtggaccgg ctgaagagcc
     3181 tgatcaaata caagggctac caggtagccc cagccgaact ggagagcatc ctgctgcaac
     3241 accccaacat cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc
     3301 ccgccgcagt cgtcgtgctg gaacacggta aaaccatgac cgagaaggag atcgtggact
     3361 atgtggccag ccaggttaca accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg
     3421 aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa gatccgcgag attctcatta
     3481 aggccaagaa gggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc
     3541 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3601 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3661 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3721 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3781 atccgtttgc gtattgggcg ctcttccgct gatctgcgca gcaccatggc ctgaaataac
     3841 ctctgaaaga ggaacttggt tagctacctt ctgaggcgga aagaaccagc tgtggaatgt
     3901 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3961 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4021 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4081 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4141 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4201 cttttttgga ggcctaggct tttgcaaaaa gctcgattct tctgacacta gcgccaccat
     4261 gatcgaacaa gacggcctcc atgctggcag tcccgcagct tgggtcgaac gcttgttcgg
     4321 gtacgactgg gcccagcaga ccatcggatg tagcgatgcg gccgtgttcc gtctaagcgc
     4381 tcaaggccgg cccgtgctgt tcgtgaagac cgacctgagc ggcgccctga acgagcttca
     4441 agacgaggct gcccgcctga gctggctggc caccaccggc gtaccctgcg ccgctgtgtt
     4501 ggatgttgtg accgaagccg gccgggactg gctgctgctg ggcgaggtcc ctggccagga
     4561 tctgctgagc agccaccttg cccccgctga gaaggtttct atcatggccg atgcaatgcg
     4621 gcgcctgcac accctggacc ccgctacctg ccccttcgac caccaggcta agcatcggat
     4681 cgagcgtgct cggacccgca tggaggccgg cctggtggac caggacgacc tggacgagga
     4741 gcatcagggc ctggcccccg ctgaactgtt cgcccgactg aaagcccgca tgccggacgg
     4801 tgaggacctg gttgtcacac acggagatgc ctgcctccct aacatcatgg tcgagaatgg
     4861 ccgcttctcc ggcttcatcg actgcggtcg cctaggagtt gccgaccgct accaggacat
     4921 cgccctggcc acccgcgaca tcgctgagga gcttggcggc gagtgggccg accgcttctt
     4981 agtcttgtac ggcatcgcag ctcccgacag ccagcgcatc gccttctacc gcttgctcga
     5041 cgagttcttt taatgatcta gaaccggtca tggccgcaat aaaatatctt tattttcatt
     5101 acatctgtgt gttggttttt tgtgtgttcg aactagatgc tgtcgaccga tgcccttgag
     5161 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     5221 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     5281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6181 aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg
     6241 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
     6301 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
     6361 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
     6421 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
     6481 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
     6541 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
     6601 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
     6661 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
     6721 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
     6781 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
     6841 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
     6901 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
     6961 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
     7021 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
     7081 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
     7141 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
     7201 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     7261 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     7321 ctgtccccag tgcaagtgca ggtgccagaa catttctct
//
