LOCUS       sequence-417164-        5371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recognition domain for in vitro MAPK-activity sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5371)
  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 5371)
  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: RC(WW)-LgBit
FEATURES             Location/Qualifiers
     source          1..5371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      22..477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(109..128)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     319..340
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        503..607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(608..676,677..1468)
                     /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(826..845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1639..2227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2128..2147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2381..2398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2413..2552
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2478..2497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2638..2660
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2654..2845)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3354..3373
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3354..3372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3434..4534
                     /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      4391..4400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4508..4531
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4592..4612
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5225..5242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5295..5313)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5309..5356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
       61 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      121 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      181 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      241 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      301 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      361 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      421 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
      481 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      541 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      601 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      661 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      721 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      781 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      841 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      901 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      961 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1021 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1081 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1141 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1201 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1261 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1321 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1381 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1441 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1501 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1741 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2341 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2401 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2461 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     2521 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     2581 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2641 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     2701 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     2761 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     2821 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     2881 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     2941 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     3001 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     3061 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     3121 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     3181 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     3241 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     3301 tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg
     3361 actcactata gggagaccac aacggtttcc ctctagattt tcacgagcac ttcaccaaca
     3421 aggaccatag attatgaaaa ctgaagaagg taaactggta atctggatta acggcgataa
     3481 aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt
     3541 caccgttgag catccggata aactggaaga gaaattccca caggttgcgg caactggcga
     3601 tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct
     3661 gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga
     3721 tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct
     3781 gatttataac aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct
     3841 ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta
     3901 cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa
     3961 gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct
     4021 ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc
     4081 tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat
     4141 cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc
     4201 caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct
     4261 ggcaaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa
     4321 agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc
     4381 acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca
     4441 gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca
     4501 gactgtcgat gaagccctga aagacgcgca gactaattcg agctcgaaca acaacactag
     4561 tggctgcggt ggtggtggcg gttccatgag cgagaatctt tattttcagg gatccatgaa
     4621 actgccgccg ggttgggaga aacgtatgag ccgcagcagc ggtcgtgtgt attattttaa
     4681 tcatatcacc aacgccagcc agtgggagcg tccgtcaggt aacagctgca gcggtagcgg
     4741 ccgcgtcttc acactcgaag atttcgttgg ggactgggaa cagacagccg cctacaacct
     4801 ggaccaagtc cttgaacagg gaggtgtgtc cagtttgctg cagaatctcg ccgtgtccgt
     4861 aactccgatc caaaggattg tccggagcgg tgaaaatgcc ctgaagatcg acatccatgt
     4921 catcatcccg tatgaaggtc tgagcgccga ccaaatggcc cagatcgaag aggtgtttaa
     4981 ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgccctatg gcacactggt
     5041 aatcgacggg gttacgccga acatgctgaa ctatttcgga cggccgtatg aaggcatcgc
     5101 cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat
     5161 cgacgagcgc ctgatcaccc ccgacggctc catgctgttc cgagtaacca tcaacagcct
     5221 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
     5281 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     5341 cttgaggggt tttttgctga aaggaggaac t
//
