LOCUS       sequence-231520-        8624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for the p38 activity reporter mKO-MK2 in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8624)
  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 8624)
  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-mKO-MK2
FEATURES             Location/Qualifiers
     source          1..8624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..658
                     /label=chicken beta-actin promoter
     intron          659..1675
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1599..1621
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1683..1702
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1733..2386
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELGEDAVAHS"
     misc_feature    3678..4227
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3822..3839)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4049..4068
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(4698..4715)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4704..4928
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4975..4991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4999..5015
                     /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(5023..5029,5030..5047,5048..5053))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5068..5089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5323..5385
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5426..5446)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5426..5444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5465..5481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5489..5505
                     /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(5513..5519,5520..5537,5538..5543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5558..5579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5696..5713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5867..6455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5947..5966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6626..7417,7418..7486))
                     /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     7249..7268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7487..7591)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7617..8072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7754..7775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7966..7985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8199..8221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8213..8230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8214..8230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8240..8259
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8240..8258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   8288..8322
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg ggggrggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     1081 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     1321 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     1381 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattca ccatggtgag
     1741 cgtgatcaag cccgagatga agatgaggta ctacatggac ggctccgtca atgggcatga
     1801 gttcacaatc gagggtgagg gcacaggcag accttacgag ggacatcagg agatgacact
     1861 gcgcgtcaca atggccaagg gcgggccaat gcctttcgcc ttcgacctgg tgtcccacgt
     1921 gttctgttac ggccacagac cttttactaa atatccagaa gagatcccag actatttcaa
     1981 gcaggccttt cctgagggcc tgtcctggga gaggtccctg gagttcgagg acggcggctc
     2041 cgcctccgtg agcgcccaca tcagcctgag gggcaacacc ttctaccaca agtccaagtt
     2101 caccggcgtg aacttccccg ccgacggccc catcatgcag aaccagagcg tggactggga
     2161 gccctccacc gagaagatca ccgccagcga cggcgtgctg aagggcgacg tgaccatgta
     2221 cctgaagctg gagggcggcg gcaaccacaa gtgccagttc aagaccacct acaaggccgc
     2281 caagaagatc ctgaagatgc ccggcagcca ctacatcagc cacaggctgg tgaggaagac
     2341 cgagggcaac atcaccgagc tgggggagga cgccgtggcc cactccctcg agatgctgtc
     2401 caactcccag ggccagagcc cgccggtgcc gttccccgcc ccggccccgc cgccgcagcc
     2461 ccccacccct gccctgccgc accccccggc gcagccgccg ccgccgcccc cgcagcagtt
     2521 cccgcagttc cacgtcaagt ccggcctgca gatcaagaag aacgccatca tcgatgacta
     2581 caaggtcacc agccaggtcc tggggctggg catcaacggc aaagttttgc agatcttcaa
     2641 caagaggacc caggagaaat tcgccctcaa aatgcttcag gactgcccca aggcccgcag
     2701 ggaggtggag ctgcactggc gggcctccca gtgcccgcac atcgtacgga tcgtggatgt
     2761 gtacgagaat ctgtacgcag ggaggaagtg cctgctgatt gtcatggaat gtttggacgg
     2821 tggagaactc tttagccgaa tccaggatcg aggagaccag gcattcacag aaagagaagc
     2881 atccgaaatc atgaagagca tcggtgaggc catccagtat ctgcattcaa tcaacattgc
     2941 ccatcgggat gtcaagcctg agaatctctt atacacctcc aaaaggccca acgccatcct
     3001 gaaactcact gactttggct ttgccaagga aaccaccagc cacaactctt tgaccactcc
     3061 ttgttataca ccgtactatg tggctccaga agtgctgggt ccagagaagt atgacaagtc
     3121 ctgtgacatg tggtccctgg gtgtcatcat gtacatcctg ctgtgtgggt atcccccctt
     3181 ctactccaac cacggccttg ccatctctcc gggcatgaag actcgcatcc gaatgggcca
     3241 gtatgaattt cccaacccag aatggtcaga agtatcagag gaagtgaaga tgctcattcg
     3301 gaatctgctg aaaacagagc ccacccagag aatgaccatc accgagttta tgaaccaccc
     3361 ttggatcatg caatcaacaa aggtccctca aaccccactg cacaccagcc gggtcctgaa
     3421 ggaggacaag gagcggtggg aggatgtcaa ggaggagatg accagtgcct tggccacaat
     3481 gcgcgttgac tacgagcaga tcaagataaa aaagattgaa gatgcatcca accctctgct
     3541 gctgaagagg cggaagaaag ctcgggccct ggaggctgcg gctctggccc actgaggcgg
     3601 ccgcggtaac aattgttaac taacttaagc tagcaacggt ttccctctag cgggatcaat
     3661 tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     3721 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     3781 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     3841 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     3901 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     3961 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4021 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     4081 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     4141 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     4201 ggttttcctt tgaaaaacac gataatacca tggtcatgaa aacatttaac atttctcaac
     4261 aagatctaga attagtagaa gtagcgacag agaagattac aatgctttat gaggataata
     4321 aacatcatgt gggagcggca attcgtacga aaacaggaga aatcatttcg gcagtacata
     4381 ttgaagcgta tataggacga gtaactgttt gtgcagaagc cattgcgatt ggtagtgcag
     4441 tttcgaatgg acaaaaggat tttgacacga ttgtagctgt tagacaccct tattctgacg
     4501 aagtagatag aagtattcga gtggtaagtc cttgtggtat gtgtagggag ttgatttcag
     4561 actatgcacc agattgtttt gtgttaatag aaatgaatgg caagttagtc aaaactacga
     4621 ttgaagaact cattccactc aaatataccc gaaattaaaa gtcgagatgc atcgatgatc
     4681 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4741 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4801 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4861 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4921 ctctatggct tctgaggcgg aaagaacctg cagcccaagc ttggcgtaat catggtcata
     4981 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatcttaa aagttttgtt
     5161 actttataga agaaattttg agtttttgtt tttttttaat aaataaataa acataaataa
     5221 attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact taatatctat
     5281 tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc aattttacgc
     5341 atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaat ctagtatacg
     5401 cgtttaatta atccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     5461 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5521 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5581 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5641 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5701 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6301 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     7621 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7681 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7741 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7801 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7861 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7921 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7981 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8041 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     8101 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     8161 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     8221 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattggggc gcgccattct
     8281 agattaaccc tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct
     8341 ctctctttct aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt
     8401 ggagctcccg tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg
     8461 accgcgtgag tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat
     8521 acgataatta tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt
     8581 tcttgttata gatatcaact agaatgctag catgggccca tctc
//
