LOCUS       sequence-172463-        6002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of GFP-JNK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6002)
  AUTHORS   Zehorai E, Seger R
  TITLE     Beta-like importins mediate the nuclear translocation of 
            mitogen-activated protein kinases.
  JOURNAL   Mol Cell Biol. 2014 Jan;34(2):259-70. doi: 10.1128/MCB.00799-13. 
            Epub 2013 Nov 11.
  PUBMED    24216760
REFERENCE   2  (bases 1 to 6002)
  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: 
            "10.1128/MCB.00799-13"; journalName: "Mol Cell Biol"; date: 
            "2014-01"; volume: "34"; issue: "2"; pages: "259-70"
COMMENT     Sequence Label: GFP-JNK1
FEATURES             Location/Qualifiers
     source          1..6002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        129..432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        433..636
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     587..607
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      675..684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(681..683,684..686,687..1397)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(681..683,684..686,687..1397)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(726..747)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(987..1006)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1334..1355
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2858..2979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2895..2914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2949..2968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2986..3441)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3123..3144)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3335..3354
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3468..3572
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3574..3931
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3574..3765
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3597..3617)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3782..3917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3966..4760
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3966..4760
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4020..4039)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4630..4649
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4948..4967)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4992..5039
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5368..5956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5857..5876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
       61 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      121 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      181 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      241 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      301 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      361 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      421 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      481 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      541 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      601 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      661 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      721 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      781 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      841 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      901 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      961 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1021 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1081 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1141 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1201 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1261 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1321 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1381 tggacgagct gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg
     1441 acagcagaag caagcgtgac aacaattttt atagtgtaga gattggagat tctacattca
     1501 cagtcctgaa acgatatcag aatttaaaac ctataggctc aggagctcaa ggaatagtat
     1561 gcgcagctta tgatgccatt cttgaaagaa atgttgcaat caagaagcta agccgaccat
     1621 ttcagaatca gactcatgcc aagcgggcct acagagagct agttcttatg aaatgtgtta
     1681 atcacaaaaa tataattggc cttttgaatg ttttcacacc acagaaatcc ctagaagaat
     1741 ttcaagatgt ttacatagtc atggagctca tggatgcaaa tctttgccaa gtgattcaga
     1801 tggagctaga tcatgaaaga atgtcctacc ttctctatca gatgctgtgt ggaatcaagc
     1861 accttcattc tgctggaatt attcatcggg acttaaagcc cagtaatata gtagtaaaat
     1921 ctgattgcac tttgaagatt cttgacttcg gtctggccag gactgcagga acgagtttta
     1981 tgatgacgcc ttatgtagtg actcgctact acagagcacc cgaggtcatc cttggcatgg
     2041 gctacaagga aaacgttgac atttggtcag ttgggtgcat catgggagaa atgatcaaag
     2101 gtggtgtttt gttcccaggt acagatcata ttgatcagtg gaataaagtt attgaacagc
     2161 ttggaacacc atgtcctgaa ttcatgaaga aactgcaacc aacagtaagg acttacgttg
     2221 aaaacagacc taaatatgct ggatatagct ttgagaaact cttccctgat gtccttttcc
     2281 cagctgactc agaacacaac aaacttaaag ccagtcaggc aagggatttg ttatccaaaa
     2341 tgctggtaat agatgcatct aaaaggatct ctgtagatga agctctccaa cacccgtaca
     2401 tcaatgtctg gtatgatcct tctgaagcag aagctccacc accaaagatc cctgacaagc
     2461 agttagatga aagggaacac acaatagaag agtggaaaga attgatatat aaggaagtta
     2521 tggacttgga ggagagaacc aagaatggag ttatacgggg gcagccctct cctttagcac
     2581 aggtgcagca gtgatcaatg gctctcagca tccatcatca tcgtcgtctg tcaatgatgt
     2641 gtcttcaatg tcaacagatc cgactttggc ctctgataca gacagcagtc tagaagcagc
     2701 agctgggcct ctgggctgct gtagatgagg atccaccgga tctagataac tgatcataat
     2761 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2821 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2881 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2941 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     3001 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3061 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3121 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3181 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3241 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3301 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3361 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3421 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3481 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3541 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3601 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3661 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3721 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3781 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3841 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3901 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3961 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4021 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4081 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4141 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4201 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4261 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4321 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4381 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4441 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4501 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4561 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4621 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4681 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4741 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4801 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4861 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4921 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4981 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5041 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5101 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5161 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5221 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5281 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5341 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5401 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5461 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5521 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5581 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5641 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5701 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5761 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5821 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5881 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5941 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6001 gc
//
