LOCUS       sequence-417118-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro RSK-Sensor for luminescent detection of RSK-activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5008)
  AUTHORS   Poti AL, Denes L, Papp K, Bato C, Banoczi Z, Remenyi A, Alexa A
  TITLE     Phosphorylation-Assisted Luciferase Complementation Assay Designed 
            to Monitor Kinase Activity and Kinase-Domain-Mediated 
            Protein-Protein Binding.
  JOURNAL   Int J Mol Sci. 2023 Oct 3;24(19):14854. doi: 10.3390/ijms241914854.
  PUBMED    37834301
REFERENCE   2  (bases 1 to 5008)
  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: "Int J Mol 
            Sci."; date: "2023-10-3"; volume: "24(19):14854. doi"; pages: " 
            10.3390/ijms241914854"
COMMENT     Sequence Label: VF-Sensor
FEATURES             Location/Qualifiers
     source          1..5008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(114..133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     324..345
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        508..612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(613..681,682..1473)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(831..850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1644..2232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2133..2152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2386..2403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2418..2557
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2483..2502)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2643..2665
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2659..2850)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3359..3378
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3359..3377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3439..4539
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      4396..4405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4513..4536
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4597..4617
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4867..4884
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4937..4955)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4951..4998
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
       61 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
      121 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
      181 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
      241 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
      301 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
      361 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
      421 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
      481 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1141 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2281 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2341 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2401 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2461 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     2521 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     2581 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2641 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     2701 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     2761 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     2821 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     2881 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     2941 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3001 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     3061 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     3121 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     3181 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     3241 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     3301 gatcatgcgc acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaatta
     3361 atacgactca ctatagggag accacaacgg tttccctcta gattttcacg agcacttcac
     3421 caacaaggac catagattat gaaaactgaa gaaggtaaac tggtaatctg gattaacggc
     3481 gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt
     3541 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact
     3601 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct
     3661 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc
     3721 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta
     3781 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg
     3841 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa
     3901 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac
     3961 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc
     4021 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca
     4081 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc
     4141 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa
     4201 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa
     4261 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt
     4321 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa
     4381 gatccacgta ttgccgccac catggaaaac gcccagaaag gtgaaatcat gccgaacatc
     4441 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt
     4501 cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc gaacaacaac
     4561 actagtggct gcggtggtgg tggcggttcc atgagcgaga atctttattt tcagggatcc
     4621 gatgtgtgca gcgtgtttga tagcgatcat agcagcccgt ttcatagcag caatgatacc
     4681 gtctttatcc aagttactct gccccatggc ccaagatctg cttctgtatc atctataagt
     4741 ttaaccaaag gcactgatga agtgcctgtc cctcctcctg ttcctccacg aagacgacca
     4801 gaatctgccc cagcagaaag cggccgcgtg accggctacc ggctgttcga ggagattctg
     4861 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     4921 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     4981 gtcttgaggg gttttttgct gaaaggag
//
