LOCUS       sequence-246084-        3196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORF encoding ERK KTR mClover flanked by Gateway sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3196)
  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 3196)
  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-ERKKTRClover
FEATURES             Location/Qualifiers
     source          1..3196
                     /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"
     primer_bind     complement(760..781)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1368..1389
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1425..1524)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1636..1655)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1647..2456
                     /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(1714..1733)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2546..3134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3035..3054
                     /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 gcaggctcca ccatgaaggg ccgaaagcct
      481 cgggacctgg aacttccact tagcccaagc ctgctgggtg gccagggacc tgaacggact
      541 ccaggatcag gaacaagctc tggtcttcag gcaccggggc cagcgctttc gcccagcaag
      601 cgtagcggtc tcgaggaccc agcgacgcca tcgaagaagc cacgtacccc atcggtgtcg
      661 tcaaggctgg agcgcctgac cctccagtcc agctttcaat tcccatccac tagtatggtg
      721 agcaagggcg aggagctgtt caccggggtt gtgcccatcc tggtcgagct ggacggcgac
      781 gtaaacggcc acaagttcag cgtccgcggc gagggcgagg gcgatgccac caacggcaag
      841 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      901 accaccttcg gctacggcgt ggcctgcttc agccgctacc ccgaccacat gaagcagcac
      961 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctctttcaag
     1021 gacgacggta cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1081 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1141 gagtacaact tcaacagcca caacgtctat atcacggccg acaagcagaa gaacggcatc
     1201 aaggctaact tcaagatccg ccacaacgtt gaggacggca gcgtgcagct cgccgaccac
     1261 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1321 agccatcagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1381 gagttcgtga ccgccgccta agcggccgca ctcgagatat ctagacccag ctttcttgta
     1441 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1501 cagtcaaaat aaaatcatta tttgccatcc agctgcagct ctggcccgtg tctcaaaatc
     1561 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     1621 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     1681 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1741 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1801 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1861 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1921 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     1981 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtc cctgcgccgg ttgcattcga
     2041 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2101 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2161 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2221 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2281 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2341 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2401 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2461 aattggttaa ttggttgtaa cattattcag attgggcccc gttccactga gcgtcagacc
     2521 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2581 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2641 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2701 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2761 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2821 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2881 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2941 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3001 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3061 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3181 cttttgctca catgtt
//
