LOCUS       sequence-417116-        6016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recognition domain for in vitro RSK-activity sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6016)
  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 6016)
  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(14-3-3-epsilon)-LgBit
FEATURES             Location/Qualifiers
     source          1..6016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      33..488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(120..139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     330..351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        514..618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(619..687,688..1479)
                     /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(837..856)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1650..2238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2139..2158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2392..2409
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2424..2563
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2489..2508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2649..2671
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2665..2856)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3365..3384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3365..3383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3445..4545
                     /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      4402..4411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4519..4542
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4603..4623
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5881..5898
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5951..5969)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5965..6012
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
       61 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      121 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      181 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      241 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      301 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
      361 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
      421 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
      481 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      541 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      601 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      661 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      721 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      781 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
      841 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      901 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      961 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1021 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1081 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1141 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     1201 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1261 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1321 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1381 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1441 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1501 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1561 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1621 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1681 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1741 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     1801 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1861 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1921 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1981 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2041 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2101 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2161 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2221 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2281 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2341 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2401 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2461 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2521 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     2581 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2641 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2701 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     2761 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     2821 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     2881 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     2941 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3001 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3061 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     3121 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     3181 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     3241 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     3301 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct cgatcccgcg
     3361 aaattaatac gactcactat agggagacca caacggtttc cctctagatt ttcacgagca
     3421 cttcaccaac aaggaccata gattatgaaa actgaagaag gtaaactggt aatctggatt
     3481 aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc
     3541 ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg
     3601 gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct
     3661 caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg
     3721 tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa
     3781 gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag
     3841 atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg
     3901 caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat
     3961 gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt
     4021 ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc
     4081 atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca
     4141 tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag
     4201 ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg
     4261 aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa
     4321 gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg
     4381 gcgaaagatc cacgtattgc cgccaccatg gaaaacgccc agaaaggtga aatcatgccg
     4441 aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc
     4501 agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaattc gagctcgaac
     4561 aacaacacta gtggctgcgg tggtggtggc ggttccatga gcgagaatct ttattttcag
     4621 ggatccatgg atgatcgaga ggatctggtg taccaggcga agctggccga gcaggctgag
     4681 cgatacgacg aaatggtgga gtcaatgaag aaagtagcag ggatggatgt ggagctgaca
     4741 gttgaagaaa gaaacctcct atctgttgca tataagaatg tgattggagc tagaagagcc
     4801 tcctggagaa taatcagcag cattgaacag aaagaagaaa acaagggagg agaagacaag
     4861 ctaaaaatga ttcgggaata tcggcaaatg gttgagactg agctaaagtt aatctgttgt
     4921 gacattctgg atgtactgga caaacacctc attccagcag ctaacactgg cgagtccaag
     4981 gttttctatt ataaaatgaa aggggactac cacaggtatc tggcagaatt tgccacagga
     5041 aacgacagga aggaggctgc ggagaacagc ctagtggctt ataaagctgc tagtgatatt
     5101 gcaatgacag aacttccacc aacgcatcct attcgcttag gtcttgctct caatttttcc
     5161 gtattctact acgaaattct taattcccct gaccgtgcct gcaggttggc aaaagcagct
     5221 tttgatgatg caattgcaga actggatacg ctgagtgaag aaagctataa ggactctaca
     5281 cttatcatgc agttgttacg tgataatctg acactatgga cttcagacat gcagggtgac
     5341 ggtgaagagc agaataaaga agcgctgcag gacgtggaag acgaaaatca gagcggccgc
     5401 gtcttcacac tcgaagattt cgttggggac tgggaacaga cagccgccta caacctggac
     5461 caagtccttg aacagggagg tgtgtccagt ttgctgcaga atctcgccgt gtccgtaact
     5521 ccgatccaaa ggattgtccg gagcggtgaa aatgccctga agatcgacat ccatgtcatc
     5581 atcccgtatg aaggtctgag cgccgaccaa atggcccaga tcgaagaggt gtttaaggtg
     5641 gtgtaccctg tggatgatca tcactttaag gtgatcctgc cctatggcac actggtaatc
     5701 gacggggtta cgccgaacat gctgaactat ttcggacggc cgtatgaagg catcgccgtg
     5761 ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac
     5821 gagcgcctga tcacccccga cggctccatg ctgttccgag taaccatcaa cagcctcgag
     5881 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     5941 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     6001 aggggttttt tgctga
//
