LOCUS       sequence-217713-        7668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for the JNK activity reporter JNK KTR-mCherry in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7668)
  AUTHORS   Miura H, Kondo Y, Matsuda M, Aoki K
  TITLE     Cell-to-Cell Heterogeneity in p38-Mediated Cross-Inhibition of JNK 
            Causes Stochastic Cell Death.
  JOURNAL   Cell Rep. 2018 Sep 4;24(10):2658-2668. doi: 
            10.1016/j.celrep.2018.08.020.
  PUBMED    30184500
REFERENCE   2  (bases 1 to 7668)
  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.celrep.2018.08"; journalName: "Cell Rep"; date: 
            "2018-09-4- 4"; volume: "24"; issue: "10"; pages: "2658-2668"
COMMENT     Sequence Label: pPBbsr-JNK KTR-mCherry
FEATURES             Location/Qualifiers
     source          1..7668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1393..1668
                     /label=chicken beta-actin promoter
     intron          1669..2677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2601..2623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             2936..3646
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTLGMDELSR"
     primer_bind     complement(3081..3099)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3349..3368
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3733..4281
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3877..3894)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4104..4123
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(4752..4769)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4758..4982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5029..5045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5053..5069
                     /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(5077..5083,5084..5101,5102..5107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5122..5143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5377..5439
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5480..5500)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5480..5498)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5519..5535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5543..5559
                     /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(5567..5573,5574..5591,5592..5597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5612..5633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5750..5767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5921..6509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6001..6020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6680..7471,7472..7540))
                     /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     7303..7322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7541..7645)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     1441 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     1501 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     1561 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     1621 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     1681 ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg
     1741 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1801 cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc
     1861 cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
     1921 ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1981 gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     2041 gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     2101 agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg
     2161 ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg
     2221 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     2281 gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc
     2341 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     2401 ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     2461 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     2521 gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg gctgccttcg
     2581 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     2641 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttgt
     2701 tgtgctgtct catcattttg gcaaagaatt gggatccgaa ttcaccatga gtaaccctaa
     2761 aatactgaag cagagcatga ccttgaacct ggccgacccg gtgggcagtc tgaagccgca
     2821 cctccgcgcc aagcgctcgg ccggtctcaa cgaagacgag tcgccatcta agaagctggc
     2881 gtcgccggag gtgtcgtcaa ggctggagcg cctgaccctc cagtccagcg gccgcatggt
     2941 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     3001 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     3061 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     3121 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     3181 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3241 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     3301 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     3361 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     3421 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     3481 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     3541 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     3601 cgagggccgc cactccactc tcggcatgga cgagctgtct agataagtcg acgggccgcg
     3661 gtaacaattg ttaactaact taagctagca acggtttccc tctagcggga tcaattccgc
     3721 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     3781 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     3841 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     3901 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     3961 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     4021 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     4081 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     4141 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     4201 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     4261 cctttgaaaa acacgataat accatggtca tgaaaacatt taacatttct caacaagatc
     4321 tagaattagt agaagtagcg acagagaaga ttacaatgct ttatgaggat aataaacatc
     4381 atgtgggagc ggcaattcgt acgaaaacag gagaaatcat ttcggcagta catattgaag
     4441 cgtatatagg acgagtaact gtttgtgcag aagccattgc gattggtagt gcagtttcga
     4501 atggacaaaa ggattttgac acgattgtag ctgttagaca cccttattct gacgaagtag
     4561 atagaagtat tcgagtggta agtccttgtg gtatgtgtag ggagttgatt tcagactatg
     4621 caccagattg ttttgtgtta atagaaatga atggcaagtt agtcaaaact acgattgaag
     4681 aactcattcc actcaaatat acccgaaatt aaaagtcgag atgcatcgat gatctagagc
     4741 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4801 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4861 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4921 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4981 ggcttctgag gcggaaagaa cctgcagccc aagcttggcg taatcatggt catagctgtt
     5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc ttaaaagttt tgttacttta
     5221 tagaagaaat tttgagtttt tgtttttttt taataaataa ataaacataa ataaattgtt
     5281 tgttgaattt attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat
     5341 taataaataa acctcgatat acagaccgat aaaacacatg cgtcaatttt acgcatgatt
     5401 atctttaacg tacgtcacaa tatgattatc tttctagggt taatctagta tacgcgttta
     5461 attaatccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5521 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5581 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5641 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5701 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5761 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5821 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5881 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5941 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6001 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6061 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6121 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6181 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6241 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6301 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6361 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6421 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6661 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6721 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6781 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6841 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6901 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6961 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7021 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7081 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7141 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7201 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7261 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7321 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7381 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7441 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7501 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7561 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7621 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
