LOCUS Exported 9085 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS LRRK2_1867_2161_L1914A_L1976Y_C2024S_C2025S_I2049Q SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9085) AUTHORS MJFF TITLE Michael J Fox Foundation plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 9085) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9085 /organism="synthetic DNA construct" /mol_type="other DNA" misc_recomb 329..1156 /label=baculovirus recombination region (lef2/ORF603) /note="contains ORF603 and part of lef2" CDS 329..453 /codon_start=3 /product="baculovirus late expression factor 2" /label=lef2 /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY" CDS complement(491..1096) /codon_start=1 /product="baculovirus ORF603 protein" /label=ORF603 /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR" CDS complement(1167..2978) /codon_start=1 /gene="uidA (or gusA) from E. coli" /product="beta-glucuronidase" /label=GUS /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW ADEHGIVVIDETAAVGFSLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK PKSAAFLLQKRWTGMNFGEKPQQGGKQ" promoter complement(2987..3302) /gene="Autographa californica p6.9" /label=p6.9 promoter /note="baculovirus late basic protein promoter" promoter 3317..3408 /gene="polh from Autographa californica" /label=polyhedrin promoter /note="promoter for the baculovirus polyhedrin gene" primer_bind 3333..3350 /label=Polyhedrin-F /note="Polyhedrin promoter, forward primer" CDS 3421..3444 /codon_start=1 /product="8xHis affinity tag" /label=8xHis /translation="HHHHHHHH" CDS 3445..3465 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3469..4122 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" CDS 4195..4215 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 5135..5152 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" misc_recomb 5189..6546 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" CDS complement(5205..6546) /codon_start=2 /product="baculovirus capsid-associated protein" /label=ORF1629 /note="required for viral replication" /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA" primer_bind complement(6638..6655) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6809..7397) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6889..6908) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7568..8428) /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 8191..8210 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8429..8533) /gene="bla" /label=AmpR promoter rep_origin 8560..9015 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8647..8666) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8857..8878 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ggaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag 61 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtcga catgctgaac 121 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat 181 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg 241 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac 301 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat 361 aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac 421 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat 481 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa 541 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac 601 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt 661 cttcgtattc cttctctttt tcatttttct cttcataaaa attaacatag ttattatcgt 721 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt 781 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat 841 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt 901 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa 961 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt 1021 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt 1081 taaaaataac agccattgta atgagacgca caaactaata tcacaaactg gaaatgtcta 1141 tcaatatata gttgctgatc agatcctcat tgtttgcctc cctgctgcgg tttttcaccg 1201 aagttcatgc cagtccagcg tttttgcagc agaaaagccg ccgacttcgg tttgcggtcg 1261 cgagtgaaga tccctttctt gttaccgcca acgcgcaata tgccttgcga ggtcgcaaaa 1321 tcggcgaaat tccatacctg ttcaccgacg acggcgctga cgcgatcaaa gacgcggtga 1381 tacatatcca gccatgcaca ctgatactct tcactccaca tgtcggtgta cattgagtgc 1441 agcccggcta acgtatccac gccgtattcg gtgatgataa tcggctgatg cagtttctcc 1501 tgccaggcca gaagttcttt ttccagtacc ttctctgccg tttccaaatc gccgctttgg 1561 acataccatc cgtaataacg gttcaggcac agcacatcaa agagatcgct gatggtatcg 1621 gtgtgagcgt cgcagaacat tacattgacg caggtgatcg gacgcgtcgg gtcgagttta 1681 cgcgttgctt ccgccagtgg cgcgaaatat tcccgtgcac cttgcggacg ggtatccggt 1741 tcgttggcaa tactccacat caccacgctt gggtggtttt tgtcacgcgc tatcagctct 1801 ttaatcgcct gtaagtgcgc ttgctgagtt tccccgttga ctgcctcttc gctgtacagt 1861 tctttcggct tgttgcccgc ttcgaaacca atgcctaaag agagcgaaaa gccgacagca 1921 gcagtttcat caatcaccac gatgccatgt tcatctgccc agtcgagcat ctcttcagcg 1981 taagggtaat gcgaggtacg gtaggagttg gccccaatcc agtccattaa tgcgtggtcg 2041 tgcaccatca gcacgttatc gaatcctttg ccacgtaagt ccgcatcttc atgacgacca 2101 aagccagtaa agtagaacgg tttgtggtta atcaggaact gttggccctt cactgccact 2161 gaccggatgc cgacgcgaag cgggtagata tcacactctg tctggctttt ggctgtgacg 2221 cacagttcat agagataacc ttcacccggt tgccagaggt gcggattcac cacttgcaaa 2281 gtcccgctag tgccttgtcc agttgcaacc acctgttgat ccgcatcacg cagttcaacg 2341 ctgacatcac cattggccac cacctgccag tcaacagacg cgtggttaca gtcttgcgcg 2401 acatgcgtca ccacggtgat atcgtccacc caggtgttcg gcgtggtgta gagcattacg 2461 ctgcgatgga ttccggcata gttaaagaaa tcatggaagt aagactgctt tttcttgccg 2521 ttttcgtcgg taatcaccat tcccggcggg atagtctgcc agttcagttc gttgttcaca 2581 caaacggtga tacgtacact tttcccggca ataacatacg gcgtgacatc ggcttcaaat 2641 ggcgtatagc cgccctgatg ctccatcact tcctgattat tgacccacac tttgccgtaa 2701 tgagtgaccg catcgaaacg cagcacgata cgctggcctg cccaaccttt cggtataaag 2761 acttcgcgct gataccagac gttgcccgca taattacgaa tatctgcatc ggcgaactga 2821 tcgttaaaac tgcctggcac agcaattgcc cggctttctt gtaacgcgct ttcccaccaa 2881 cgctgatcaa ttccacagtt ttcgcgatcc agactgaatg cccacaggcc gtcgagtttt 2941 ttgatttcac gggttggggt ttctacagga cggaccatat ttataggtgt aatttatgta 3001 gctgtaattt ttaccttatt aatatttttt acgctttgca ttcgacgact gaactcccaa 3061 atatatgttt aactcgtctt ggtcgtttga atttttgttg ctgtgtttcc taatattttc 3121 catcacctta aatatgttat tgtaatcctc aatgttgaac ttgcaattgg acacggcata 3181 gttttccata gtcgtgtaaa acatggtatt ggctgcattg taatacatcc gactgagcgg 3241 gtacggatct atgtgtttga gcagcctgtt caaaaactct gcatcgtcgc aaaacggaat 3301 ttcgtcgaca gatctgatca tggagataat taaaatgata accatctcgc aaataaataa 3361 gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat aggatctatg 3421 caccaccatc accaccatca ccacgagaac ctttatttcc aaggatcaat ggcaccgatc 3481 ctggggtatt ggaagatcaa gggcctagtg cagcccactc gtctgctcct ggagtaccta 3541 gaagagaagt atgaggagca cctttatgag cgggacgagg gtgacaagtg gcgcaataag 3601 aaattcgagc taggcttgga attccccaat ctgccttatt acatcgatgg cgacgttaag 3661 ttgactcagt ccatggccat catccgctac atcgccgaca agcacaatat gctaggcggg 3721 tgcccgaagg agcgagctga gatctctatg ttggaagggg ccgtgctgga tatccgctac 3781 ggcgtgagcc gcattgcata ctcaaaggat ttcgagaccc tcaaggttga cttcctgtca 3841 aagctgcctg aaatgctaaa gatgttcgaa gaccgtctct gtcataagac ctacctgaac 3901 ggggaccacg tgacacaccc ggactttatg ttgtatgacg ctctggacgt cgtgctttat 3961 atggacccta tgtgcctgga cgcttttccc aaactagtgt gtttcaagaa acgtatcgaa 4021 gctatccctc agattgataa gtacctgaag tcctcaaagt acatcgcctg gccccttcaa 4081 ggttggcaag ccacatttgg cggtggcgac catcctccca agtcagacct agtgcctagg 4141 cacaaccaga ccagcctata taagaaagcc gggtcagctg ccgctgtgct ggaggagaac 4201 ctgtatttcc agggcggatc caacatcatg ctcaacaacg atgagctcga atttgaacaa 4261 gctccagaat tcctgctcgg cgacggctct ttcggctccg tataccgtgc tgcttatgaa 4321 ggcgaagaag tcgccgtaaa gatcttcaac aagcacacct cggctcgtct gctccgtcag 4381 gaactggtcg ttctgtgtca tctgcaccat ccgagcctca tctccctgct ggccgctggc 4441 atccgtcctc gtatgctggt gatggagctg gcatccaagg gctctctcga ccgcctgctg 4501 cagcaggaca aggcttctct gacccgtact ctgcagcatc gtatcgctta ccatgtagct 4561 gacggcctgc gttatcttca ttctgcaatg atcatttatc gcgacctgaa acctcataat 4621 gtactgctgt ttactctgta cccgaacgct gccatcatcg ctaagattgc agactatggt 4681 attgcgcagt actcctcccg catgggcatt aaaacaagcg aaggtacgcc gggtttccgt 4741 gctccagaag tggctcgtgg caacgttcag tacaaccagc aagctgacgt ttactcgttc 4801 ggtctgctcc tgtacgatat cctgactacc ggcggccgta tcgtagaagg tctgaagttc 4861 cctaacgaat tcgacgaact ggaaatccag ggcaagctgc cagacccggt taaagaatac 4921 ggttgcgctc cgtggccgat ggtagagaag ctcatcaaac agtgccttaa agaaaaccct 4981 caggaacgtc caacctctgc tcaagtgttc gacatcctta acagcgctga actcgtctgt 5041 ctgactcgcc gtattctcct gcctaagaat gttatcgtgg aatgtatggt tgctactcac 5101 cataactgat gataagcttt agtaagcact cgagcaccac caccaccacc actaacctag 5161 gtagctgagc gcatgcaagc tgatccgggt tattagtaca tttattaagc gctagattct 5221 gtgcgttgtt gatttacaga caattgttgt acgtatttta ataattcatt aaatttataa 5281 tctttagggt ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt atatctgaat 5341 ttaaatatta aatcctcaat agatttgtaa aataggtttc gattagtttc aaacaagggt 5401 tgtttttccg aaccgatggc tggactatct aatggatttt cgctcaacgc cacaaaactt 5461 gccaaatctt gtagcagcaa tctagctttg tcgatattcg tttgtgtttt gttttgtaat 5521 aaaggttcga cgtcgttcaa aatattatgc gcttttgtat ttctttcatc actgtcgtta 5581 gtgtacaatt gactcgacgt aaacacgtta aatagagctt ggacatattt aacatcgggc 5641 gtgttagctt tattaggccg attatcgtcg tcgtcccaac cctcgtcgtt agaagttgct 5701 tccgaagacg attttgccat agccacacga cgcctattaa ttgtgtcggc taacacgtcc 5761 gcgatcaaat ttgtagttga gctttttgga attatttctg attgcgggcg tttttgggcg 5821 ggtttcaatc taactgtgcc cgattttaat tcagacaaca cgttagaaag cgatggtgca 5881 ggcggtggta acatttcaga cggcaaatct actaatggcg gcggtggtgg agctgatgat 5941 aaatctacca tcggtggagg cgcaggcggg gctggcggcg gaggcggagg cggaggtggt 6001 ggcggtgatg cagacggcgg tttaggctca aatgtctctt taggcaacac agtcggcacc 6061 tcaactattg tactggtttc gggcgccgtt tttggtttga ccggtctgag acgagtgcga 6121 tttttttcgt ttctaatagc ttccaacaat tgttgtctgt cgtctaaagg tgcagcgggt 6181 tgaggttccg tcggcattgg tggagcgggc ggcaattcag acatcgatgg tggtggtggt 6241 ggtggaggcg ctggaatgtt aggcacggga gaaggtggtg gcggcggtgc cgccggtata 6301 atttgttctg gtttagtttg ttcgcgcacg attgtgggca ccggcgcagg cgccgctggc 6361 tgcacaacgg aaggtcgtct gcttcgaggc agcgcttggg gtggtggcaa ttcaatatta 6421 taattggaat acaaatcgta aaaatctgct ataagcattg taatttcgct atcgtttacc 6481 gtgccgatat ttaacaaccg ctcaatgtaa gcaattgtat tgtaaagaga ttgtctcaag 6541 ctcggatcga tcccgcacgc cgataacaag ccttttcatt tttactacag cattgtagtg 6601 gcgagacact tcgctgtcgt cgaggtttaa acgcttcctc gctcactgac tcgctgcgct 6661 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 6721 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 6781 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 6841 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 6901 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 6961 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 7021 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 7081 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 7141 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 7201 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 7261 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 7321 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 7381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 7441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 7501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 7561 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 7621 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 7681 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 7741 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 7801 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 7861 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 7921 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 7981 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 8041 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 8101 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 8161 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 8221 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 8281 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 8341 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 8401 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 8461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 8521 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat 8581 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 8641 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 8701 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 8761 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 8821 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 8881 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 8941 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 9001 taacgtttac aatttcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 9061 tgcgggcctc ttcgctatta cgcca //