LOCUS       sequence-256102-        6135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Myc tagged human ERK2(K52R) for purification by GST 
            affinity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6135)
  AUTHORS   Smolko CM, Janes KA
  TITLE     An ultrasensitive fiveplex activity assay for cellular kinases.
  JOURNAL   Sci Rep. 2019 Dec 19;9(1):19409. doi: 10.1038/s41598-019-55998-8.
  PUBMED    31857650
REFERENCE   2  (bases 1 to 6135)
  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: "Sci Rep."; 
            date: "2019-12-19"; volume: "9(1):19409. doi"; pages: " 
            10.1038/s41598-019-55998-8"
COMMENT     Sequence Label: pGEX-4T-1-3xMyc-ERK2(K52R)
FEATURES             Location/Qualifiers
     source          1..6135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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)."
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             198..725
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(218..235)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(218..234)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(227..249)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(827..832,833..848,849..855)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    863..879
                     /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)."
     CDS             902..1555
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1513..1535
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1562..1579
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1583..1672
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     regulatory      1683..1692
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2720..2755
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     primer_bind     complement(2829..2851)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2996..3014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3082..3186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3187..3255,3256..4047)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3405..3424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4218..4806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4707..4726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4960..4977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5050..5127
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5128..6135,1..75)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5147..5166)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
      241 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      301 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      361 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
      421 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
      481 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
      541 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
      601 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
      661 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
      721 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
      781 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca
      841 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt
      901 catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
      961 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
     1021 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
     1081 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
     1141 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
     1201 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
     1261 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
     1321 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
     1381 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
     1441 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
     1501 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga
     1561 tctggttccg cgtggatcta tggagcagaa actcatctct gaagaggatc tggagcagaa
     1621 actcatctct gaagaggatc tggagcagaa actcatctct gaagaggatc tgaagcttgg
     1681 atccatggcg gcggcggcgg cgggcccgga gatggtccgc gggcaggtgt tcgacgtggg
     1741 gccgcgctac actaatctct cgtacatcgg agaaggcgcc tacggcatgg tttgttctgc
     1801 ttatgataat ctcaacaaag ttcgagttgc tatcaggaaa atcagtcctt ttgagcacca
     1861 gacctactgt cagagaaccc tgagagagat aaaaatccta ctgcgcttca gacatgagaa
     1921 catcatcggc atcaatgaca tcatccgggc accaaccatt gagcagatga aagatgtata
     1981 tatagtacag gacctcatgg agacagatct ttacaagctc ttgaagacac agcacctcag
     2041 caatgatcat atctgctatt ttctttatca gatcctgaga ggattaaagt atatacattc
     2101 agctaatgtt ctgcaccgtg acctcaagcc ttccaacctc ctgctgaaca ccacttgtga
     2161 tctcaagatc tgtgactttg gccttgcccg tgttgcagat ccagaccatg atcatacagg
     2221 gttcttgaca gagtatgtag ccacgcgttg gtacagagct ccagaaatta tgttgaattc
     2281 caagggttat accaagtcca ttgatatttg gtctgtgggc tgcatcctgg cagagatgct
     2341 atccaacagg cctatcttcc caggaaagca ttaccttgac cagctgaatc acatcctggg
     2401 tattcttgga tctccatcac aggaagatct gaattgtata ataaatttaa aagctagaaa
     2461 ctatttgctt tctctcccgc acaaaaataa ggtgccgtgg aacagattgt tcccaaacgc
     2521 tgactccaaa gctctggatt tactggataa aatgttgaca tttaaccctc acaagaggat
     2581 tgaagttgaa caggctctgg cccacccgta cctggagcag tattatgacc caagtgatga
     2641 gcccattgct gaagcaccat tcaagtttga catggagctg gacgacttac ctaaggagaa
     2701 gctcaaagaa ctcatttttg aagagactgc tcgattccag ccaggataca gatcttaagt
     2761 cgactcgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac
     2821 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2881 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     2941 gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc
     3001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3421 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3721 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4981 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5041 ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     5101 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     5161 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     5221 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     5281 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     5341 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     5401 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     5461 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     5521 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     5581 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     5641 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     5701 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     5761 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     5821 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     5881 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     5941 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     6001 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     6061 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     6121 cccaatacgc aaacc
//
