LOCUS       sequence-363826-        7896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA tagged ERK5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7896)
  AUTHORS   Buschbeck M, Eickhoff J, Sommer MN, Ullrich A
  TITLE     Phosphotyrosine-specific phosphatase PTP-SL regulates the ERK5 
            signaling pathway.
  JOURNAL   J Biol Chem. 2002 Aug 16;277(33):29503-9. Epub 2002 May 31.
  PUBMED    12042304
REFERENCE   2  (bases 1 to 7896)
  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: "J Biol 
            Chem."; date: "2002-08-16"; volume: "277(33)"; pages: "29503-9. Epub
            2002 May 31"
COMMENT     Sequence Label: pcDNA3-HA-ERK5
FEATURES             Location/Qualifiers
     source          1..7896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             672..698
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(3204..3222)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3205..3222)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3242..3259)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3248..3472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3518..3946
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3605..3624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3815..3836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3955..3975)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3960..4289
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4140..4275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4202..4221
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4356..5150
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4410..4429)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5020..5039
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5324..5445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5361..5380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5415..5434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5494..5510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5494..5510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5507..5529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5518..5534
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5542..5548,5549..5566,5567..5572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5587..5608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5725..5742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5907..6495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5987..6006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6666..7457,7458..7526))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7289..7308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7527..7631)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7706..7725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ccgccaccat gtatgatgtt cctgattatg ctagcctccc gggaggatcc gccgagcctc
      721 tgaaggagga agacggcgag gacggctctg cggagccccc cgggcccgtg aaggccgaac
      781 ccgcccacac cgctgcctct gtagcggcca agaacctggc cctgcttaaa gcccgctcct
      841 tcgatgtgac ctttgacgtg ggcgacgagt acgagatcat cgagaccata ggcaacgggg
      901 cctatggagt ggtgtcctcc gcccgccgcc gcctcaccgg ccagcaggtg gccatcaaga
      961 agatccctaa tgctttcgat gtggtgacca atgccaagcg gaccctcagg gagctgaaga
     1021 tcctcaagca cttcaaacac gacaacatca tcgccatcaa ggacatcctg aggcccaccg
     1081 tgccctatgg cgaattcaaa tctgtctacg tggtcctgga cctgatggaa agcgacctgc
     1141 accagatcat ccactcctca cagcccctca cactggaaca cgtgcgctac ttcctgtacc
     1201 aactgctgcg gggcctgaag tacatgcact cggctcaggt catccaccgt gacctgaagc
     1261 cctccaacct attggtgaat gagaactgtg agctcaagat tggtgacttt ggtatggctc
     1321 gtggcctgtg cacctcgccc gctgaacatc agtacttcat gactgagtat gtggccacgc
     1381 gctggtaccg tgcgcccgag ctcatgctct ctttgcatga gtatacacag gctattgacc
     1441 tctggtctgt gggctgcatc tttggtgaga tgctggcccg gcgccagctc ttcccaggca
     1501 aaaactatgt acaccagcta cagctcatca tgatggtgct gggtacccca tcaccagccg
     1561 tgattcaggc tgtgggggct gagagggtgc gggcctatat ccagagcttg ccaccacgcc
     1621 agcctgtgcc ctgggagaca gtgtacccag gtgccgaccg ccaggcccta tcactgctgg
     1681 gtcgcatgct gcgttttgag cccagcgctc gcatctcagc agctgctgcc cttcgccacc
     1741 ctttcctggc caagtaccat gatcctgatg atgagcctga ctgtgccccg ccctttgact
     1801 ttgcctttga ccgcgaagcc ctcactcggg agcgcattaa ggaggccatt gtggctgaaa
     1861 ttgaggactt ccatgcaagg cgtgagggca tccgccaaca gatccgcttc cagccttctc
     1921 tacagcctgt gactagtgag cctggctgtc cagatgttga aatgcccagt ccctgggctc
     1981 ccagtgggga ctgtgccatg gagtctccac caccagcccc gccaccatgc cccggccctg
     2041 cacctgacac cattgatctg accctgcagc cacctccacc agtcagtgag cctgccccac
     2101 caaagaaaga tggtgccatc tcagacaata ctaaggctgc ccttaaagct gccctgctca
     2161 agtctttgag gagccggctc agagatggcc ccagcgcacc cctggaggct cctgagcctc
     2221 ggaagccggt gacagcccag gagcgccagc gggagcggga ggagaagcgg cggaggcggc
     2281 aagaacgagc caaggagcgg gagaaacggc ggcaggagcg ggagcgaaag gaacgggggg
     2341 ctggggcctc tgggggcccc tccactgacc ccttggctgg actagtgctc agtgacaatg
     2401 acagaagcct gttggaacgc tggactcgaa tggcccggcc cgcagcccca gccctcacct
     2461 ctgtgccggc ccctgcccca gcgccaacgc caaccccaac cccagtccaa cctaccagtc
     2521 ctcctcctgg ccctgtagcc cagcccactg gcccgcaacc acaatctgcg ggctctacct
     2581 ctggccctgt accccagcct gcctgcccac cccctggccc tgcaccccac cccactggcc
     2641 ctcctgggcc catccctgtc cccgcgccac cccagattgc cacctccacc agcctcctgg
     2701 ctgcccagtc acttgtgcca ccccctgggc tgcctggctc cagcacccca ggagttttgc
     2761 cttacttccc acctggcctg ccgcccccag acgccggggg agcccctcag tcttccatgt
     2821 cagagtcacc tgatgtcaac cttgtgaccc agcagctatc taagtcacag gtggaggacc
     2881 ccctgccccc tgtgttctca ggcacaccaa agggcagtgg ggctggctac ggtgttggct
     2941 ttgacctgga ggaattctta aaccagtctt tcgacatggg cgtggctgat gggccacagg
     3001 atggccaggc agattcagcc tctctctcag cctccctgct tgctgactgg ctcgaaggcc
     3061 atggcatgaa ccctgccgat attgagtccc tgcagcgtga gatccagatg gactccccaa
     3121 tgctgctggc tgacctgcct gacctccagg acccctgagg cccccagcct gtgccttgct
     3181 gccacagttc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     3241 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3481 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4261 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4321 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4381 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4441 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4501 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4561 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4621 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4681 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4741 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4801 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4861 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4921 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4981 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5041 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5101 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5161 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5221 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5281 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5341 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5401 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5461 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5521 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5581 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5641 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5701 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5761 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5821 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccagcaaaa ggccaggaac
     5881 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5941 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6001 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6061 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6121 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6181 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6241 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6301 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6361 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6421 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6481 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6541 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6601 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6661 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6721 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6781 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6841 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6901 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6961 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7021 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7081 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7141 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7201 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7261 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7321 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7381 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7441 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7501 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7561 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7621 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7681 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7741 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7801 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     7861 cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
