LOCUS       sequence-176198-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes negative-control C-terminal (substrate) fragment of 
            bimolecular JNK activity reporter (bimJNKAR); cytosol targeted; use 
            in conjunction with pcDNA3-Cerulean-FHA1-NES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  AUTHORS   Depry C, Mehta S, Li R, Zhang J
  TITLE     Visualization of Compartmentalized Kinase Activity Dynamics Using 
            Adaptable BimKARs.
  JOURNAL   Chem Biol. 2015 Nov 19;22(11):1470-9. doi: 
            10.1016/j.chembiol.2015.10.004. Epub 2015 Nov 5.
  PUBMED    26548610
REFERENCE   2  (bases 1 to 6314)
  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.chembiol.2015.10.004"; journalName: "Chem Biol"; date: 
            "2015-11-19- 19"; volume: "22"; issue: "11"; pages: "1470-9"
COMMENT     Sequence Label: pcDNA3-JNKsub(TA)DD-YPet-NES
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             22..54
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(58..66,67..81)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(484..503)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             901..927
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(946..964)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(947..964)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(984..1001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    990..1214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1260..1688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1347..1366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1557..1578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1697..1717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1702..2031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1882..2017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1944..1963
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2098..2892
                     /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(2152..2171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2762..2781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3066..3187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3103..3122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3157..3176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3236..3252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3236..3252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3249..3271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3260..3276
                     /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(3284..3290,3291..3308,3309..3314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3329..3350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3467..3484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3638..4226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3718..3737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4397..5188,5189..5257))
                     /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     5020..5039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5258..5362)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5437..5456)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5628..6007
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6008..6211
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6162..6182
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6256..6275
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6256..6274
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6294..6303
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
       61 ctgtacgacg atgacgataa ggatcccatg gtcgacagtg tcaaggcccc cgaggatgaa
      121 ggcaacccac tgctcgagca gctcgagaag aagggcggca ccggcggcag cgagctcatg
      181 gtgagcaaag gcgaagagct gttcaccggc gtggtgccca tcctggtgga gctggacggc
      241 gacgtgaacg gccacaagtt cagcgtgagc ggcgagggcg agggcgacgc cacctacggc
      301 aagctgaccc tgaagctgct gtgcaccacc ggcaagctgc ccgtgccctg gcccaccctg
      361 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccggt accccgacca catgaagcag
      421 cacgacttct tcaagagcgc catgcccgag ggctacgtgc aggagcggac catcttcttc
      481 aaggacgacg gcaactacaa gacccgggcc gaggtgaagt tcgagggcga caccctggtg
      541 aaccggatcg agctgaaggg catcgacttc aaggaggaca gcaacatcct gggccacaag
      601 ctggagtaca actacaacag ccacaacgtg tacatcaccg ccgacaagca gaagaacggc
      661 atcaaggcca acttcaagat ccggcacaac atcgaggacg gcggcgtgca gctggccgac
      721 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga taaccactac
      781 ctgagctacc agagcgccct gttcaaggac cccaacgaga agcgggacca catggtgctg
      841 ctggagttcc tgaccgccgc cggcatcacc gagggtatga atgaattgta caaagaattc
      901 ctgccccccc tggagcgcct gaccctgtaa tctagagggc cctattctat agtgtcacct
      961 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1021 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1081 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1141 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     1201 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1261 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1321 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1381 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1441 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1501 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1561 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1621 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1681 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     1741 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     1801 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     1861 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     1921 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     1981 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2041 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     2101 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2161 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2221 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     2281 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     2341 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     2401 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     2461 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     2521 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     2581 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     2641 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     2701 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     2761 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     2821 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     2881 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     2941 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3001 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3061 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3181 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3241 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3301 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3361 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3421 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3541 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4201 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4321 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4381 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4441 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4501 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4561 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4621 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4681 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4741 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4801 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4861 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4921 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4981 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5041 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5101 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5161 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5221 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5281 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5341 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5401 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     5461 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     5521 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     5581 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     5641 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5701 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     5761 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     5821 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5881 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     5941 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6001 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     6061 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     6121 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     6181 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     6241 ctggcttatc gaaattaata cgactcacta tagggagacc caagcttgcg gccgccacca
     6301 tggtgcgggg ttct
//
