LOCUS       sequence-356966-        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 T183AY185F
FEATURES             Location/Qualifiers
     source          1..5764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(630..632,633..635,636..1346)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(630..632,633..635,636..1346)
                     /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(675..696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(936..955)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1283..1304
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2400..2435
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     polyA_signal    2569..2690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2606..2625)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2660..2679
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2697..3152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3046..3065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3179..3283
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3285..3642
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3285..3476
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3308..3328)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3493..3628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3677..4471
                     /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             3677..4471
                     /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(3731..3750)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4341..4360
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4659..4678)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4703..4750
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5079..5667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5568..5587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      661 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      721 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      781 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      841 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      901 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      961 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1021 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1081 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1141 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1201 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1261 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1321 ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagcggcg gcggcggcgg
     1381 cgggcccgga gatggtccgc gggcaggtgt ttgacgtagg gccgcgctac accaacctct
     1441 cgtacatcgg agaaggcgcc tacggcatgg tttgctctgc ttatgataat ctcaacaaag
     1501 ttcgagttgc tatcaagaaa atcagtcctt ttgagcacca gacctactgt caaagaaccc
     1561 taagagagat aaaaatctta ctgcgcttca gacatgagaa catcattggc atcaatgaca
     1621 tcatccgggc accaaccatt gagcaaatga aagatgtata tatagtacag gacctcatgg
     1681 agacggacct ttacaagctc ttgaagacac agcacctcag caatgaccac atctgctatt
     1741 ttctttatca gatcctgaga gggctaaagt atatccattc agctaacgtt ctgcaccgtg
     1801 acctcaagcc ttccaacctc ctgctgaaca ccacttgtga tctcaagatc tgtgactttg
     1861 gccttgcccg tgttgcagat ccagatcatg atcacacagg gttcttggca gaatttgtcg
     1921 cgacgcgttg gtacagagct ccagaaatta tgttaaattc caagggttat accaagtcca
     1981 ttgatatttg gtctgtgggc tgcatcctgg cagagatgct atccaacagg cctatcttcc
     2041 caggaaagca ttaccttgac cagctgaatc acatcctggg tattcttgga tctccatcac
     2101 aggaagatct gaattgtata ataaatttaa aagctagaaa ctatttgctt tctctcccgc
     2161 acaaaaataa ggtgccatgg aacaggttgt tcccaaatgc tgactccaaa gctctggatt
     2221 tactggataa aatgttgaca tttaaccctc acaagaggat tgaagttgaa caggctctgg
     2281 cccacccata cctggagcag tattatgacc caagtgatga gcccattgct gaagcgccat
     2341 tcaagtttga catggagttg gacgacttac ctaaggagaa gctcaaagaa ctcatttttg
     2401 aagagactgc tagattccag ccaggataca gatcttagag gatccaccgg atctagataa
     2461 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2521 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2581 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2641 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     2701 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2761 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2821 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2881 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3121 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3181 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3241 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3301 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3421 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3481 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3541 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3601 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3661 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3721 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3781 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3841 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3901 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3961 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4021 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4081 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4141 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4201 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4261 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4321 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4381 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4441 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4501 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4561 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4621 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4681 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4741 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4801 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4861 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4921 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5701 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5761 aacc
//
