LOCUS       sequence-329215-        6259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for His6-Delta-MEK1C218.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6259)
  AUTHORS   Lai KY, Galan SRG, Zeng Y, Zhou TH, He C, Raj R, Riedl J, Liu S, 
            Chooi KP, Garg N, Zeng M, Jones LH, Hutchings GJ, Mohammed S, Nair 
            SK, Chen J, Davis BG, van der Donk WA
  TITLE     LanCLs add glutathione to dehydroamino acids generated at 
            phosphorylated sites in the proteome.
  JOURNAL   Cell. 2021 Apr 27. pii: S0092-8674(21)00436-0. doi: 
            10.1016/j.cell.2021.04.001.
  PUBMED    33932340
REFERENCE   2  (bases 1 to 6259)
  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: "Cell. 2021 
            Apr 27. pii: S0092-8674(21)00436-0. doi: 
            10.1016/j.cell.2021.04.001."
COMMENT     Sequence Label: Delta-1-60MEK1C218
FEATURES             Location/Qualifiers
     source          1..6259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     157..176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        157..175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    176..200
                     /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             1213..1230
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1280..1297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1350..1368)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1364..1411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1448..1903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1535..1554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1745..1766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1995..2810)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1995..2810)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2718..2737
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2802..2821
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2932..3520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3421..3440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3674..3691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3706..3845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3771..3790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3931..3953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3947..4138)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4947..6029)
                     /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     5991..6010
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6030..6107)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
       61 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
      121 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat
      181 tgtgagcgga taacaattcc cctctagaaa taattttgtt taaccttaag aaggagatat
      241 actatggaac tgaaagatga cgatttcgaa aaaatcagtg aactgggcgc cggtaacggc
      301 ggtgtggttt ttaaagtcag ccataaaccg tctggtctgg tgatggcacg taaactgatt
      361 cacctggaaa tcaaaccggc tattcgtaac cagattatcc gcgaactgca agtgctgcat
      421 gaatgcaatt ctccgtatat tgttggcttt tatggtgcgt tctacagtga cggcgaaatt
      481 tccatctgta tggaacacat ggacggcggt agcctggatc aggttctgaa aaaagcaggc
      541 cgtatcccgg aacaaattct gggtaaagtc tctattgctg tgatcaaagg cctgacgtac
      601 ctgcgtgaaa aacataaaat catgcaccgc gatgtgaaac cgtcaaacat cctggttaat
      661 tcgcgcggtg aaattaaact gtgcgacttt ggcgttagcg gtcagctgat tgattgtatg
      721 gcgaactctt tcgtcggcac ccgtagttat atgtccccgg aacgcctgca gggtacgcat
      781 tactcagtgc aatcggatat ctggtcaatg ggcctgtcgc tggttgaaat ggccgttggt
      841 cgttatccga ttccgccgcc ggacgccaaa gaactggaac tgatgtttgg ctctcaggtt
      901 gaaggtgatg cggccgaaac cccgccgcgt ccgcgtaccc cgggtcgtcc gctgagctct
      961 tatggtatgg acagccgtcc gccgatggca atcttcgaac tgctggatta cattgtgaat
     1021 gaaccgccgc caaaactgcc gagcggcgtg tttagcctgg aatttcagga cttcgtcaac
     1081 aaatgtctga tcaaaaatcc ggcagaacgt gctgatctga aacaactgat ggtccacgct
     1141 tttattaaac gctccgacgc ggaagaagtg gatttcgccg gttggctgtc ttcgaccatt
     1201 ggcctgaatg cgcaccatca tcaccaccat tgaggatccg aattcgagct ccgtcgacaa
     1261 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     1321 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1381 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     1441 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1501 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1561 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1621 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1681 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1741 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1801 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1861 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     1921 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1981 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     2041 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     2101 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     2161 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     2221 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     2281 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     2341 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     2401 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     2461 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     2521 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     2581 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     2641 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     2701 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     2761 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     2821 gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga
     2881 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3061 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3241 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3301 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3361 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3421 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3481 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3541 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3601 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3661 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3721 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3781 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3841 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4021 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4081 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4141 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4201 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4261 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4321 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4381 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4441 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4501 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4561 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     4621 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     4681 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     4741 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     4801 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     4861 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     4921 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4981 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     5041 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     5101 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     5161 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     5221 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     5281 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     5341 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     5401 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     5461 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     5521 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     5581 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     5641 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     5701 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     5761 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     5821 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     5881 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     5941 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     6001 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     6061 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     6121 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     6181 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     6241 ggggcctgcc accataccc
//
