LOCUS       sequence-172461-        5820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of GFP-p38beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  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 5820)
  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-p38beta
FEATURES             Location/Qualifiers
     source          1..5820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        215..518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        519..722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     673..693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      761..770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(767..769,770..772,773..1483)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(767..769,770..772,773..1483)
                     /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(812..833)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1073..1092)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1420..1441
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2762..2883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2799..2818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2853..2872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2890..3345)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3027..3048)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3239..3258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3372..3476
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3478..3835
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3478..3669
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3501..3521)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3686..3821
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3870..4664
                     /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             3870..4664
                     /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(3924..3943)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4534..4553
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4852..4871)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4896..4943
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5272..5820,1..40)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5761..5780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
       61 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      121 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
      181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      481 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      541 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      601 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      661 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      721 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg
      781 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      841 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      901 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      961 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1021 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1081 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1141 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1201 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1261 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1321 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1381 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1441 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctcg
     1501 agctcaagct tcgaattcca tgtcgggccc tcgcgccggc ttctaccggc aggagctgaa
     1561 caagaccgtg tgggaggtgc cgcagcggct gcaggggctg cgcccggtgg gctccggcgc
     1621 ctacggctcc gtctgttcgg cctacgacgc ccggctgcgc cagaaggtgg cggtgaagaa
     1681 gctgtcgcgc cccttccagt cgctgatcca cgcgcgcaga acgtaccggg agctgcggct
     1741 gctcaagcac ctgaagcacg agaacgtcat cgggcttctg gacgtcttca cgccggccac
     1801 gtccatcgag gacttcagcg aagtgtactt ggtgaccacc ctgatgggcg ccgacctgaa
     1861 caacatcgtc aagtgccagg cgctgagcga cgagcacgtt caattcctgg tttaccagct
     1921 gctgcgcggg ctgaagtaca tccactcggc cgggatcatc caccgggacc tgaagcccag
     1981 caacgtggct gtgaacgagg actgtgagct caggatcctg gatttcgggc tggcgcgcca
     2041 ggcggacgag gagatgaccg gctatgtggc cacgcgctgg taccgggcac ctgagatcat
     2101 gctcaactgg atgcattaca accaaacagt ggatatctgg tccgtgggct gcatcatggc
     2161 tgagctgctc cagggcaagg ccctcttccc gggaagcgac tacattgacc agctgaagcg
     2221 catcatggaa gtggtgggca cacccagccc tgaggttctg gcaaaaatct cctcggaaca
     2281 cgcccggaca tatatccagt ccctgccccc catgccccag aaggacctga gcagcatctt
     2341 ccgtggagcc aaccccctgg ccatagacct ccttggaagg atgctggtgc tggacagtga
     2401 ccagagggtc agtgcagctg aggcactggc ccacgcctac ttcagccagt accacgaccc
     2461 cgaggatgag ccagaggccg agccatatga tgagagcgtt gaggccaagg agcgcacgct
     2521 ggaggagtgg aaggagctca cttaccagga agtcctcagc ttcaagcccc cagagccacc
     2581 gaagccacct ggcagcctgg agattgagca ggtcgacggt accgcgggcc cgggatccac
     2641 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2701 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2761 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2881 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2941 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3001 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3061 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3121 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3181 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3241 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3301 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3481 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3541 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3661 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3721 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3781 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3841 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3901 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3961 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4021 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4081 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4141 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4201 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4261 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4321 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4381 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4441 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4501 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4561 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4621 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4681 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4741 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4801 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4861 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4921 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4981 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5041 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5101 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
//
