LOCUS       sequence-227427-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Rat ERK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  AUTHORS   Horgan AM, Stork PJ
  TITLE     Examining the mechanism of Erk nuclear translocation using green 
            fluorescent protein.
  JOURNAL   Exp Cell Res. 2003 May 1;285(2):208-20.
  PUBMED    12706116
REFERENCE   2  (bases 1 to 5752)
  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"; journalName: "Exp Cell 
            Res."; date: "2003-05-1"; volume: "285(2)"; pages: "208-20"
COMMENT     Sequence Label: GFP-ERK2 del174-177
FEATURES             Location/Qualifiers
     source          1..5752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      629..638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(635..637,638..640,641..1351)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(635..637,638..640,641..1351)
                     /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(680..701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(941..960)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1288..1309
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2393..2428
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     polyA_signal    2562..2683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2599..2618)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2653..2672
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2690..3145)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2827..2848)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3039..3058
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3172..3276
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3278..3635
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3278..3469
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3301..3321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3486..3621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3670..4464
                     /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             3670..4464
                     /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(3724..3743)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4334..4353
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4652..4671)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4696..4743
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5072..5660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5561..5580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
      661 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      721 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      781 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      841 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      901 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      961 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1021 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1081 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1141 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1201 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1261 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1321 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag cggcggcggc
     1381 ggcggcgggc ccggagatgg tccgcgggca ggtgtttgac gtagggccgc gctacaccaa
     1441 cctctcgtac atcggagaag gcgcctacgg catggtttgc tctgcttatg ataatctcaa
     1501 caaagttcga gttgctatca agaaaatcag tccttttgag caccagacct actgtcaaag
     1561 aaccctaaga gagataaaaa tcttactgcg cttcagacat gagaacatca ttggcatcaa
     1621 tgacatcatc cgggcaccaa ccattgagca aatgaaagat gtatatatag tacaggacct
     1681 catggagacg gacctttaca agctcttgaa gacacagcac ctcagcaatg accacatctg
     1741 ctattttctt tatcagatcc tgagagggct aaagtatatc cattcagcta acgttctgca
     1801 ccgtgacctc aagccttcca acctcctgct gaacaccact tgtgatctca agatctgtga
     1861 ctttggcctt gcccgtgttg cagatcatac agggttcttg acagagtacg tagccacacg
     1921 ttggtacaga gctccagaaa ttatgttgaa ttccaagggt tataccaagt ccattgatat
     1981 ttggtctgtg ggctgcatcc tggcagagat gctatccaac aggcctatct tcccaggaaa
     2041 gcattacctt gaccagctga atcacatcct gggtattctt ggatctccat cacaggaaga
     2101 tctgaattgt ataataaatt taaaagctag aaactatttg ctttctctcc cgcacaaaaa
     2161 taaggtgcca tggaacaggt tgttcccaaa tgctgactcc aaagctctgg atttactgga
     2221 taaaatgttg acatttaacc ctcacaagag gattgaagtt gaacaggctc tggcccaccc
     2281 atacctggag cagtattatg acccaagtga tgagcccatt gctgaagcgc cattcaagtt
     2341 tgacatggag ttggacgact tacctaagga gaagctcaaa gaactcattt ttgaagagac
     2401 tgctagattc cagccaggat acagatctta gaggatccac cggatctaga taactgatca
     2461 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2521 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2581 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2641 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     2701 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2761 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2821 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2881 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2941 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3001 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3061 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3121 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3181 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3241 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3301 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3481 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3541 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3601 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3661 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3721 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3781 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3841 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3901 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3961 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4021 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4081 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4141 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4201 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4261 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4321 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4381 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4441 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4501 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4561 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4621 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4681 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4741 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4801 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4861 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4921 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4981 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5701 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg ga
//
