LOCUS       sequence-100426-        7390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based biosensor for monitoring JNK activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7390)
  AUTHORS   Fosbrink M, Aye-Han NN, Cheong R, Levchenko A, Zhang J
  TITLE     Visualization of JNK activity dynamics with a genetically encoded 
            fluorescent biosensor.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Mar 23;107(12):5459-64. doi: 
            10.1073/pnas.0909671107. Epub 2010 Mar 8.
  PUBMED    20212108
REFERENCE   2  (bases 1 to 7390)
  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: 
            "10.1073/pnas.0909671107"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2010-03-23- 23"; volume: "107"; issue: "12"; pages: "5459-64"
COMMENT     Sequence Label: pcDNA3-JNKAR1
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(914..916,917..919,920..1432)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             join(2162..2164,2165..2167,2168..2878)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSSLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(2942..2960)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2943..2960)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2980..2997)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2986..3210
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3256..3684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3343..3362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3553..3574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3693..3713)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3698..4028
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3879..4014
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3941..3960
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4095..4889
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4149..4168)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4759..4778
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5063..5184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5100..5119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5154..5173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5233..5249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5233..5249)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5246..5268)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5257..5273
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5281..5287,5288..5305,5306..5311))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5326..5347
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5464..5481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5635..6223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5715..5734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6394..7185,7186..7254))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7017..7036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7255..7359)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      961 ggtcgagctg gacggcgacg taaacggcca caggttcagc gtgtccggcg agggcgaggg
     1021 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1081 gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc
     1141 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1201 gcgtaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1261 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1321 catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga
     1381 caagcagaag aacggcatca aggcccactt caagatccgc cacaacatcg aggacggcag
     1441 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1501 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1561 cgatcacatg gtcctgctgg agttcgtgac cgccgcccgc atgcataagt tttctcaaga
     1621 acagatcggc gaaaacattg tgtgcagggt catttgtacc acgggtcaaa ttcccatccg
     1681 agatttgtca gctgatattt cacaagtgct taaggaaaaa cgatccataa agaaagtttg
     1741 gacatttggt agaaacccag cctgtgacta tcatttagga aacatttcaa gactgtcaaa
     1801 taagcatttc caaatactac taggagaaga cggtaacctt ttattgaatg acatttccac
     1861 taatgggacc tggttaaatg ggcaaaaagt cgagaagaac agcaatcagt tactgtctca
     1921 aggtgatgaa ataaccgttg gtgtaggcgt ggaatcagat attttatctc tggtcatttt
     1981 cataaacgac aaatttaagc agtgcctcga gcagaacaaa gttgatcgct ctgcaggtaa
     2041 gccaggcagc ggcgagggca gcaccaaggg cctggtcgac agtgtcaaga cccccgagga
     2101 tgaaggcaac ccactgctcg agcagctcga gaagaagggc ggcaccggcg gcagcgagct
     2161 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2221 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2281 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2341 cctcgtgacc accttcggct acggcctgat gtgcttcgcc cgctaccccg accacatgaa
     2401 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2461 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2521 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2581 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2641 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2701 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2761 ctacctgagc taccagtcct ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2821 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2881 agaattctgc agatatccat cacactggcg gccgctcgag catgcatcta gagggcccta
     2941 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3001 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3061 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3121 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3181 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3241 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3301 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3361 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     3421 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3481 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3541 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3601 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     3661 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3721 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3781 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3841 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3901 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3961 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4021 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4081 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4141 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4201 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4261 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4321 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4381 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4441 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4501 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4561 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4621 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4681 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4741 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4801 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4861 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4921 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4981 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5041 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5101 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5161 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5221 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5281 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5341 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5401 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5461 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5521 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5581 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5641 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5701 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5761 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5821 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5881 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5941 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6001 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6061 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6241 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6301 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6361 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6421 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6481 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6541 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6601 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6661 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6721 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6781 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6841 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6901 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6961 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7021 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7081 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7141 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7201 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7321 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7381 acctgacgtc
//
