LOCUS       sequence-227429-        5764 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 5764)
  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 5764)
  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 K52R
FEATURES             Location/Qualifiers
     source          1..5764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(645..647,648..650,651..1361)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(645..647,648..650,651..1361)
                     /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(690..711)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(951..970)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1298..1319
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2415..2450
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     polyA_signal    2584..2705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2621..2640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2675..2694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2712..3167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2849..2870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3061..3080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3194..3298
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3300..3657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3300..3491
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3323..3343)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3508..3643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3692..4486
                     /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             3692..4486
                     /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(3746..3765)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4356..4375
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4674..4693)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4718..4765
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5094..5682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5583..5602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
      661 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      721 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      781 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      841 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      901 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      961 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1021 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1081 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1141 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1201 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1261 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1321 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
     1381 cggcggcggc ggcggcgggc ccggagatgg tccgcgggca ggtgtttgac gtagggccgc
     1441 gctacaccaa cctctcgtac atcggagaag gcgcctacgg catggtttgc tctgcttatg
     1501 ataatctcaa caaagttcga gtcgcgatca ggaaaatcag tccttttgag caccagacct
     1561 actgtcaaag aaccctaaga gagataaaaa tcttactgcg cttcagacat gagaacatca
     1621 ttggcatcaa tgacatcatc cgggcaccaa ccattgagca aatgaaagat gtatatatag
     1681 tacaggacct catggagacg gacctttaca agctcttgaa gacacagcac ctcagcaatg
     1741 accacatctg ctattttctt tatcagatcc tgagagggct aaagtatatc cattcagcta
     1801 acgttctgca ccgtgacctc aagccttcca acctcctgct gaacaccact tgtgatctca
     1861 agatctgtga ctttggcctt gcccgtgttg cagatccaga tcatgatcac acagggttct
     1921 tgacagagta cgtagccaca cgttggtaca gagctccaga aattatgttg aattccaagg
     1981 gttataccaa gtccattgat atttggtctg tgggctgcat cctggcagag atgctatcca
     2041 acaggcctat cttcccagga aagcattacc ttgaccagct gaatcacatc ctgggtattc
     2101 ttggatctcc atcacaggaa gatctgaatt gtataataaa tttaaaagct agaaactatt
     2161 tgctttctct cccgcacaaa aataaggtgc catggaacag gttgttccca aatgctgact
     2221 ccaaagctct ggatttactg gataaaatgt tgacatttaa ccctcacaag aggattgaag
     2281 ttgaacaggc tctggcccac ccatacctgg agcagtatta tgacccaagt gatgagccca
     2341 ttgctgaagc gccattcaag tttgacatgg agttggacga cttacctaag gagaagctca
     2401 aagaactcat ttttgaagag actgctagat tccagccagg atacagatct tagaggatcc
     2461 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2521 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2581 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2701 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2761 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2821 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2881 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2941 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3001 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3061 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3121 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3181 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3241 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3301 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3361 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3541 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3601 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3661 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3721 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3781 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3841 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3901 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3961 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4021 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4081 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4141 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4201 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4261 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4321 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4381 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4441 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4501 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4561 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4621 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4681 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4741 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4801 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4861 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4921 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4981 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5041 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5221 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5461 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5761 cctg
//
