LOCUS Exported 6259 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression plasmid for His6-Delta-MEK1C218. ACCESSION . VERSION . KEYWORDS Delta-1-60MEK1C218 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6259 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /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 //