LOCUS       sequence-274434-        7031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human LRRK2 WD40 domain (residues 2142–2527) was expressed in Sf9 
            insect cells via baculovirus infection and purified to homogeneity..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1817889116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-01-29- 29"; volume: "116"; issue: "5"; pages: 
            "1579-1584"
COMMENT     Sequence Label: pFastBac1-LRRK2 WD40
FEATURES             Location/Qualifiers
     source          1..7031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(587..655,656..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"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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"
                     /label=Tn7L
                     /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
//
