LOCUS       sequence-91625-e        3154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORF encoding PKA KTR mClover flanked by gateway sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3154)
  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 3154)
  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-PKAKTRClover
FEATURES             Location/Qualifiers
     source          1..3154
                     /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(718..739)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1326..1347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1383..1482)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1594..1613)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1605..2414
                     /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(1672..1691)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2504..3092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2993..3012
                     /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 ccatggagat gccagaggaa
      481 cccgccaaca gtgggcattc actacccccg gtttatattt acagtcccga gtatgtcagc
      541 atctgcgact cccttgtgaa ggtccccaaa cgggccagta tggtcaacga agacgaggcg
      601 ccatctagaa ggaaggcgtc gggtcaggtg tcgtcaaggc tggagcgcct gaccctccag
      661 tccagcacta gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      721 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga gggcgagggc
      781 gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      841 ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag ccgctacccc
      901 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      961 cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt gaagttcgag
     1021 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1081 atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat cacggccgac
     1141 aagcagaaga acggcatcaa ggctaacttc aagatccgcc acaacgttga ggacggcagc
     1201 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1261 cccgacaacc actacctgag ccatcagtcc gccctgagca aagaccccaa cgagaagcgc
     1321 gatcacatgg tcctgctgga gttcgtgacc gccgcctaag cggccgcact cgagatatct
     1381 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1441 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct
     1501 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     1561 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     1621 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1681 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1741 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1801 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1861 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1921 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgtccc
     1981 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2041 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2101 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     2161 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2221 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2281 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2341 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2401 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt
     2461 tccactgagc gtcagacccg gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2581 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2641 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2701 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2761 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2821 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2881 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2941 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3001 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3061 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3121 tcctggcctt ttgctggcct tttgctcaca tgtt
//
