LOCUS Exported 7031 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Human LRRK2 WD40 domain (residues 2142–2527) was expressed in Sf9 insect cells via baculovirus infection and purified to homogeneity.. ACCESSION . VERSION . KEYWORDS pFastBac1-LRRK2 WD40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7031) AUTHORS Zhang P, Fan Y, Ru H, Wang L, Magupalli VG, Taylor SS, Alessi DR, Wu H TITLE Crystal structure of the WD40 domain dimer of LRRK2. JOURNAL Proc Natl Acad Sci U S A. 2019 Jan 29;116(5):1579-1584. doi: 10.1073/pnas.1817889116. Epub 2019 Jan 11. PUBMED 30635421 REFERENCE 2 (bases 1 to 7031) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7031 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 2..456 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(89..108) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 298..319 /label=F1ori-F /note="F1 origin, forward primer" promoter 482..586 /gene="bla" /label=AmpR promoter CDS 587..1447 /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 complement(805..824) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1618..2206 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2107..2126 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2360..2377 /label=L4440 /note="L4440 vector, forward primer" mobile_element 2511..2735 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" promoter complement(3505..3533) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" promoter 3885..3976 /gene="polh from Autographa californica" /label=polyhedrin promoter /note="promoter for the baculovirus polyhedrin gene" primer_bind 3901..3918 /label=Polyhedrin-F /note="Polyhedrin promoter, forward primer" regulatory 4019..4028 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4037..4054 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 4061..5158 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW YAVRTAVINAASGRQTVDEALKDAQT" regulatory 5015..5024 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 5132..5155 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 5186..5206 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" polyA_signal 6521..6655 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6558..6577) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6612..6631 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" mobile_element 6684..6849 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (left end of the Tn7 transposon)" ORIGIN 1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg 301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt 481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt 2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg 2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct 2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga 2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag 2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt 2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga 2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac 2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg 2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg 2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca 2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact 3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc 3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta 3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct 3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg 3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg 3301 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc 3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg 3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt 3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt 3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc 3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag 3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt 3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa 3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc 3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta 3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat 3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatctgc 4021 caccatgggc agcagccatc atcatcatca tcacggtacc aaaactgaag aaggtaaact 4081 ggtaatctgg attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt 4141 cgagaaagat accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt 4201 cccacaggtt gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt 4261 tggtggctac gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga 4321 caagctgtat ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc 4381 gatcgctgtt gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa 4441 aacctgggaa gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct 4501 gatgttcaac ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta 4561 tgcgttcaag tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg 4621 cgcgaaagcg ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga 4681 caccgattac tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa 4741 cggcccgtgg gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact 4801 gccgaccttc aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa 4861 cgccgccagt ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga 4921 tgaaggtctg gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta 4981 cgaggaagag ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg 5041 tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt 5101 gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactgg 5161 taccgattac gatatcccaa cgaccgaaaa cctttacttc cagggcggat ccagacgcat 5221 tttattacct aaaaacgtaa ttgttgaatg catggttgct acacatcaca acagcaggaa 5281 tgcaagcatt tggctgggct gtgggcacac cgacagagga cagctctcat ttcttgactt 5341 aaatactgaa ggatacactt ctgaggaagt tgctgatagt agaatattgt gcttagcctt 5401 ggtgcatctt cctgttgaaa aggaaagctg gattgtgtct gggacacagt ctggtactct 5461 cctggtcatc aataccgaag atgggaaaaa gagacatacc ctagaaaaga tgactgattc 5521 tgtcacttgt ttgtattgca attccttttc caagcaaagc aaacaaaaaa attttctttt 5581 ggttggaacc gctgatggca agttagcaat ttttgaagat aagactgtta agcttaaagg 5641 agctgctcct ttgaagatac taaatatagg aaatgtcagt actccattga tgtgtttgag 5701 tgaatccaca aattcaacgg aaagaaatgt aatgtgggga ggatgtggca caaagatttt 5761 ctccttttct aatgatttca ccattcagaa actcattgag acaagaacaa gccaactgtt 5821 ttcttatgca gctttcagtg attccaacat cataacagtg gtggtagaca ctgctctcta 5881 tattgctaag caaaatagcc ctgttgtgga agtgtgggat aagaaaactg aaaaactctg 5941 tggactaata gactgcgtgc actttttaag ggaggtaacg gtaaaagaaa acaaggaatc 6001 aaaacacaaa atgtcttatt ctgggagagt gaaaaccctc tgccttcaga agaacactgc 6061 tctttggata ggaactggag gaggccatat tttactcctg gatctttcaa ctcgtcgact 6121 tatacgtgta atttacaact tttgtaattc ggtcagagtc atgatgacag cacagctagg 6181 aagccttaaa aatgtcatgc tggtattggg ctacaaccgg aaaaatactg aaggtacaca 6241 aaagcagaaa gagatacaat cttgcttgac cgtttgggac atcaatcttc cacatgaagt 6301 gcaaaattta gaaaaacaca ttgaagtgag aaaagaatta gctgaaaaaa tgagacgaac 6361 atctgttgag tgactcgagg catgcggtac caagcttgtc gagaagtact agaggatcat 6421 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 6481 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 6541 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 6601 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc 6661 actgcttgag cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt 6721 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact 6781 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt 6841 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc 6901 catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg 6961 tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt tgcagcctga 7021 atggcgaatg g //