LOCUS       sequence-227422-        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
FEATURES             Location/Qualifiers
     source          1..5764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      602..611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(608..610,611..613,614..1324)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(608..610,611..613,614..1324)
                     /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(653..674)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(914..933)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1261..1282
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2378..2413
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     polyA_signal    2547..2668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2584..2603)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2638..2657
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2675..3130)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2812..2833)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3024..3043
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3157..3261
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3263..3620
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3263..3454
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3286..3306)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3471..3606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3655..4449
                     /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             3655..4449
                     /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(3709..3728)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4319..4338
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4637..4656)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4681..4728
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5057..5645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5546..5565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt
      601 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      661 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      721 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      781 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
      841 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      901 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      961 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1021 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1081 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1141 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1201 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1261 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1321 caagtccgga ctcagatctc gagcggcggc ggcggcggcg ggcccggaga tggtccgcgg
     1381 gcaggtgttt gacgtagggc cgcgctacac caacctctcg tacatcggag aaggcgccta
     1441 cggcatggtt tgctctgctt atgataatct caacaaagtt cgagttgcta tcaagaaaat
     1501 cagtcctttt gagcaccaga cctactgtca aagaacccta agagagataa aaatcttact
     1561 gcgcttcaga catgagaaca tcattggcat caatgacatc atccgggcac caaccattga
     1621 gcaaatgaaa gatgtatata tagtacagga cctcatggag acggaccttt acaagctctt
     1681 gaagacacag cacctcagca atgaccacat ctgctatttt ctttatcaga tcctgagagg
     1741 gctaaagtat atccattcag ctaacgttct gcaccgtgac ctcaagcctt ccaacctcct
     1801 gctgaacacc acttgtgatc tcaagatctg tgactttggc cttgcccgtg ttgcagatcc
     1861 agatcatgat cacacagggt tcttgacaga gtacgtagcc acacgttggt acagagctcc
     1921 agaaattatg ttgaattcca agggttatac caagtccatt gatatttggt ctgtgggctg
     1981 catcctggca gagatgctat ccaacaggcc tatcttccca ggaaagcatt accttgacca
     2041 gctgaatcac atcctgggta ttcttggatc tccatcacag gaagatctga attgtataat
     2101 aaatttaaaa gctagaaact atttgctttc tctcccgcac aaaaataagg tgccatggaa
     2161 caggttgttc ccaaatgctg actccaaagc tctggattta ctggataaaa tgttgacatt
     2221 taaccctcac aagaggattg aagttgaaca ggctctggcc cacccatacc tggagcagta
     2281 ttatgaccca agtgatgagc ccattgctga agcgccattc aagtttgaca tggagttgga
     2341 cgacttacct aaggagaagc tcaaagaact catttttgaa gagactgcta gattccagcc
     2401 aggatacaga tcttagagga tccaccggat ctagataact gatcataatc agccatacca
     2461 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2521 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2581 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2641 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     2701 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2761 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2821 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2881 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2941 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3001 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3061 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3121 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3181 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3241 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3301 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3361 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3421 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3481 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3541 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3601 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3661 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3721 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3781 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3841 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3901 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3961 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4021 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4081 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4141 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4201 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4261 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4321 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4381 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4441 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4501 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4561 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4621 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4681 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4741 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4801 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4861 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5701 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     5761 agtt
//
