LOCUS Exported 9383 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEMB018 / LRRK2_BC_1500_2148 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9383) AUTHORS MJFF TITLE Michael J Fox Foundation plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 9383) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9383 /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..3462 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 5433..5450 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" misc_recomb 5487..6844 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" CDS complement(5503..6844) /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(6936..6953) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7107..7695) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7187..7206) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7866..8726) /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 8489..8508 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8727..8831) /gene="bla" /label=AmpR promoter rep_origin 8858..9313 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8945..8964) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9155..9176 /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 ccacttggtg ccccgcggat ccttacgtaa gaccattatc 3481 aacgaaagcc tgaacttcaa aattcgcgac cagctggtgg tcggacagct gatccccgac 3541 tgctacgtgg aacttgagaa gatcatcctg agcgagcgca agaatgtgcc tatcgagttt 3601 ccagtgatcg atcgcaaacg tctcctccag ctcgtgcgtg agaatcagct gcagctcgac 3661 gagaacgagc tgcctcatgc tgtgcacttc cttaacgagt caggtgtcct cctgcacttc 3721 caggacccgg ctctccagct gagtgacctc tatttcgtgg agccaaagtg gctctgtaaa 3781 attatggctc aaatcctcac tgtcaaggta gagggctgtc caaagcaccc aaagggcatc 3841 atctcgcgtc gcgacgtgga aaagtttctc tctaagaagc gcaagtttcc caagaactac 3901 atgagccagt actttaaact gttggagaag ttccagatcg ctctgcctat cggcgaagag 3961 tacctgttgg ttccttcctc cctgtccgat caccgccccg tgattgagct gcctcactgc 4021 gagaacagcg agattatcat tcgcctatac gagatgccgt actttccgat gggcttctgg 4081 tcaaggctca tcaatcgtct gcttgaaatc tccccatata tgctgagcgg tcgtgaacgc 4141 gctctgaggc caaaccggat gtactggcgc caggggattt acttgaactg gtcccctgag 4201 gcttattgtc tggtgggctc ggaagtcctg gataaccacc ccgaatcatt cctgaagatc 4261 accgttcctt cctgccgtaa gggttgcatt ctattgggac aggtggtgga tcatatcgat 4321 tccctgatgg aggagtggtt ccctggcctg ttggagatcg acatctgtgg cgagggtgag 4381 accctgctga agaagtgggc actctacagt ttcaatgacg gcgaagaaca ccagaagatt 4441 ctgctcgacg acctcatgaa gaaggccgag gagggcgacc tgctcgtgaa ccccgaccag 4501 cctaggctga ccatccccat ctcccagatt gcacccgacc ttatcctcgc tgacctgcct 4561 cgcaacatca tgctgaacaa tgacgagctg gagtttgaac aagcacccga gttcctgcta 4621 ggcgacggct ctttcggttc agtttaccgt gctgcttacg agggtgaaga ggttgccgtg 4681 aagattttta acaagcacac ttcgctccgt ctgctgcgtc aggagcttgt ggttctctgc 4741 cacctgcacc acccttccct gatctccctg ctggccgccg gcattaggcc tcgcatgcta 4801 gtgatggaac tggccagtaa gggttcattg gaccgcctgc tgcagcagga caaggctagc 4861 ctgacccgta cgctccagca ccgtatcgcc ctgcatgtgg ccgacggact gcgctacttg 4921 cacagtgcca tgatcattta tcgcgatctg aagcctcata atgttcttct gttcacgctc 4981 tacccaaacg cagctatcat tgccaaaatt gccgattacg gcatagccca gtactgctgt 5041 cgtatgggta tcaagactag cgagggtaca cctgggttcc gcgctcccga agtcgcacgc 5101 gggaacgtga tctacaacca gcaggccgac gtgtactcgt ttggcctgct cctgtatgac 5161 attctcacta ccggaggccg gattgtggag ggcctgaaat ttcccaacga gttcgacgag 5221 ctggagatcc agggcaagct gcctgacccc gtgaaggagt acggttgtgc accctggcct 5281 atggtggaga agctcattaa gcagtgtctg aaggagaacc ctcaggaacg ccccacaagc 5341 gcccaggtgt tcgacattct aaactctgca gagttagtgt gtctgactag gcgtatctta 5401 ctcccgaagt aaagcttgcg gccgcactcg agcaccacca ccaccaccac taacctaggt 5461 agctgagcgc atgcaagctg atccgggtta ttagtacatt tattaagcgc tagattctgt 5521 gcgttgttga tttacagaca attgttgtac gtattttaat aattcattaa atttataatc 5581 tttagggtgg tatgttagag cgaaaatcaa atgattttca gcgtctttat atctgaattt 5641 aaatattaaa tcctcaatag atttgtaaaa taggtttcga ttagtttcaa acaagggttg 5701 tttttccgaa ccgatggctg gactatctaa tggattttcg ctcaacgcca caaaacttgc 5761 caaatcttgt agcagcaatc tagctttgtc gatattcgtt tgtgttttgt tttgtaataa 5821 aggttcgacg tcgttcaaaa tattatgcgc ttttgtattt ctttcatcac tgtcgttagt 5881 gtacaattga ctcgacgtaa acacgttaaa tagagcttgg acatatttaa catcgggcgt 5941 gttagcttta ttaggccgat tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc 6001 cgaagacgat tttgccatag ccacacgacg cctattaatt gtgtcggcta acacgtccgc 6061 gatcaaattt gtagttgagc tttttggaat tatttctgat tgcgggcgtt tttgggcggg 6121 tttcaatcta actgtgcccg attttaattc agacaacacg ttagaaagcg atggtgcagg 6181 cggtggtaac atttcagacg gcaaatctac taatggcggc ggtggtggag ctgatgataa 6241 atctaccatc ggtggaggcg caggcggggc tggcggcgga ggcggaggcg gaggtggtgg 6301 cggtgatgca gacggcggtt taggctcaaa tgtctcttta ggcaacacag tcggcacctc 6361 aactattgta ctggtttcgg gcgccgtttt tggtttgacc ggtctgagac gagtgcgatt 6421 tttttcgttt ctaatagctt ccaacaattg ttgtctgtcg tctaaaggtg cagcgggttg 6481 aggttccgtc ggcattggtg gagcgggcgg caattcagac atcgatggtg gtggtggtgg 6541 tggaggcgct ggaatgttag gcacgggaga aggtggtggc ggcggtgccg ccggtataat 6601 ttgttctggt ttagtttgtt cgcgcacgat tgtgggcacc ggcgcaggcg ccgctggctg 6661 cacaacggaa ggtcgtctgc ttcgaggcag cgcttggggt ggtggcaatt caatattata 6721 attggaatac aaatcgtaaa aatctgctat aagcattgta atttcgctat cgtttaccgt 6781 gccgatattt aacaaccgct caatgtaagc aattgtattg taaagagatt gtctcaagct 6841 cggatcgatc ccgcacgccg ataacaagcc ttttcatttt tactacagca ttgtagtggc 6901 gagacacttc gctgtcgtcg aggtttaaac gcttcctcgc tcactgactc gctgcgctcg 6961 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 7021 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 7081 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 7141 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 7201 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 7261 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 7321 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 7381 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 7441 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 7501 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 7561 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 7621 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 7681 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 7741 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 7801 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 7861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 7921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 7981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 8041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 8101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 8161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 8221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 8281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 8341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 8401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 8461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 8521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 8581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 8641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 8701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 8761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 8821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta 8881 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 8941 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 9001 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 9061 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 9121 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 9181 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 9241 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta 9301 acgtttacaa tttcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 9361 cgggcctctt cgctattacg cca //