LOCUS       sequence-417151-        4963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro p38-Sensor for luminescent detection of p38-activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4963)
  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 4963)
  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: FENEF-Sensor
FEATURES             Location/Qualifiers
     source          1..4963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      32..487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(119..138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     329..350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        513..617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(618..686,687..1478)
                     /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(836..855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1649..2237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2138..2157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2391..2408
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2423..2562
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2488..2507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2648..2670
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2664..2855)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3364..3383
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3364..3382
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3444..4544
                     /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      4401..4410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4518..4541
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4602..4622
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4827..4844
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4897..4915)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4911..4958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
       61 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      121 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      181 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      241 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      301 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      361 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      421 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      481 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      541 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      601 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      661 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      721 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      781 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      841 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
      901 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      961 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1021 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1081 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1141 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1201 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1261 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1321 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1381 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1441 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1981 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2281 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2341 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2401 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2461 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2521 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2581 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2641 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2701 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     2761 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     2821 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     2881 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     2941 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     3001 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     3061 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     3121 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     3181 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     3241 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     3301 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga
     3361 aattaatacg actcactata gggagaccac aacggtttcc ctctagattt tcacgagcac
     3421 ttcaccaaca aggaccatag attatgaaaa ctgaagaagg taaactggta atctggatta
     3481 acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg
     3541 gaattaaagt caccgttgag catccggata aactggaaga gaaattccca caggttgcgg
     3601 caactggcga tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc
     3661 aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt
     3721 ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag
     3781 cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga
     3841 tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc
     3901 aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg
     3961 aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc
     4021 tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca
     4081 tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat
     4141 ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg
     4201 gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga
     4261 acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag
     4321 cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg
     4381 cgaaagatcc acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa atcatgccga
     4441 acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca
     4501 gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg agctcgaaca
     4561 acaacactag tggctgcggt ggtggtggcg gttccatgag cgagaatctt tattttcagg
     4621 gatccgctag cgcagctcct gatgctcgtg cagtgattcc accgcatatg ggcagtggta
     4681 gcagtggcgg tagcagcggc tctggtagcg tcgacgtacc tcgtaccccg gtgagcggcc
     4741 aggcgaaccc gtttgaaaac gaatttaaaa aagcgagcag cggccgcgtg accggctacc
     4801 ggctgttcga ggagattctg ctcgagcacc accaccacca ccactgagat ccggctgcta
     4861 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     4921 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaa
//
