LOCUS       sequence-91626-e        3250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORF encoding JNK KTR mRuby2 flanked by Gateway sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3250)
  AUTHORS   Regot S, Hughey JJ, Bajar BT, Carrasco S, Covert MW
  TITLE     High-sensitivity measurements of multiple kinase activities in live 
            single cells.
  JOURNAL   Cell. 2014 Jun 19;157(7):1724-34. doi: 10.1016/j.cell.2014.04.039.
  PUBMED    24949979
REFERENCE   2  (bases 1 to 3250)
  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"; doi: 
            "10.1016/j.cell.2014.04"; journalName: "Cell"; date: "2014-06-19- 
            19"; volume: "157"; issue: "7"; pages: "1724-34"
COMMENT     Sequence Label: pENTR-JNKKTRmRuby2
FEATURES             Location/Qualifiers
     source          1..3250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    486..538
                     /label=MCS
                     /note="multiple cloning site"
     CDS             745..1452
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     protein_bind    complement(1479..1578)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1690..1709)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1701..2510
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1768..1787)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2600..3188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3089..3108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
      541 cggccggtcg ccaccatgag taaccctaag atcctaaaac agagcatgac cttgaacctg
      601 gccgacccgg tgggcagtct gaagccgcac ctccgcgcca agcgctcggc cggtctcaac
      661 gaagacgagt cgccatctaa gaagctggcg tcgccggagg tgtcgtcaag gctggagcgc
      721 ctgaccctcc agtccagcac tagtgtgtct aagggcgaag agctgatcaa ggaaaatatg
      781 cgtatgaagg tggtcatgga aggttcggtc aacggccacc aattcaaatg cacaggtgaa
      841 ggagaaggca atccgtacat gggaactcaa accatgagga tcaaagtcat cgagggagga
      901 cccctgccat ttgcctttga cattcttgcc acgtcgttca tgtatggcag ccgtactttt
      961 atcaagtacc cgaaaggcat tcctgatttc tttaaacagt cctttcctga gggttttact
     1021 tgggaaagag ttacgagata cgaagatggt ggagtcgtca ccgtcatgca ggacaccagc
     1081 cttgaggatg gctgtctcgt ttaccacgtc caagtcagag gggtaaactt tccctccaat
     1141 ggtcccgtga tgcagaagaa gaccaagggt tgggagccta atacagagat gatgtatcca
     1201 gcagatggtg gtctgagggg atacactcat atggcactga aagttgatgg tggtggccat
     1261 ctgtcttgct ctttcgtaac aacttacagg tcaaaaaaga ccgtcgggaa catcaagatg
     1321 cccggtatcc atgccgttga tcaccgcctg gaaaggttag aggaaagtga caatgaaatg
     1381 ttcgtagtac aacgcgaaca cgcagttgcc aagttcgccg ggcttggtgg tgggatggac
     1441 gagctgtaca agtaagcggc cgcactcgag atatctagac ccagctttct tgtacaaagt
     1501 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1561 aaataaaatc attatttgcc atccagctgc agctctggcc cgtgtctcaa aatctctgat
     1621 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1681 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     1741 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     1801 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     1861 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     1921 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     1981 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     2041 caggtgaaaa tattgttgat gcgctggcag tgtccctgcg ccggttgcat tcgattcctg
     2101 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     2161 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2221 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     2281 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     2341 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     2401 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2461 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     2521 ttaattggtt gtaacattat tcagattggg ccccgttcca ctgagcgtca gacccggtag
     2581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3241 ctcacatgtt
//
