LOCUS       sequence-52205-e        8594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8594)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8594)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pEMB018 / LRRK2_BC_1847_2232
FEATURES             Location/Qualifiers
     source          1..8594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             4644..4661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_recomb     4698..6055
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4714..6055)
                     /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(6147..6164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6318..6906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6398..6417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7077..7868,7869..7937))
                     /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     7700..7719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7938..8042)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8069..8524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8156..8175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8366..8387
                     /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 ccaggctgac catccccatc
     3481 tcccagattg cacccgacct tatcctcgct gacctgcctc gcaacatcat gctgaacaat
     3541 gacgagctgg agtttgaaca agcacccgag ttcctgctag gcgacggctc tttcggttca
     3601 gtttaccgtg ctgcttacga gggtgaagag gttgccgtga agatttttaa caagcacact
     3661 tcgctccgtc tgctgcgtca ggagcttgtg gttctctgcc acctgcacca cccttccctg
     3721 atctccctgc tggccgccgg cattaggcct cgcatgctag tgatggaact ggccagtaag
     3781 ggttcattgg accgcctgct gcagcaggac aaggctagcc tgacccgtac gctccagcac
     3841 cgtatcgccc tgcatgtggc cgacggactg cgctacttgc acagtgccat gatcatttat
     3901 cgcgatctga agcctcataa tgttcttctg ttcacgctct acccaaacgc agctatcatt
     3961 gccaaaattg ccgattacgg catagcccag tactgctgtc gtatgggtat caagactagc
     4021 gagggtacac ctgggttccg cgctcccgaa gtcgcacgcg ggaacgtgat ctacaaccag
     4081 caggccgacg tgtactcgtt tggcctgctc ctgtatgaca ttctcactac cggaggccgg
     4141 attgtggagg gcctgaaatt tcccaacgag ttcgacgagc tggagatcca gggcaagctg
     4201 cctgaccccg tgaaggagta cggttgtgca ccctggccta tggtggagaa gctcattaag
     4261 cagtgtctga aggagaaccc tcaggaacgc cccacaagcg cccaggtgtt cgacattcta
     4321 aactctgcag agttagtgtg tctgactagg cgtatcttac tcccgaagaa cgtgatcgtt
     4381 gagtgcatgg tggcaacgca ccacaactcg cgtaacgcta gcatctggtt aggatgtggg
     4441 cataccgatc gtggccagct gagtttcctc gacctcaaca ccgaggggta tactagcgag
     4501 gaggtcgctg attcacgcat tctgtgcctc gcccttgtcc atctgcctgt tgagaaagaa
     4561 tcgtggattg ttagtggcac ccagagcggg accctcctcg tgatcaatac cgaagacggg
     4621 taaagcttgc ggccgcactc gagcaccacc accaccacca ctaacctagg tagctgagcg
     4681 catgcaagct gatccgggtt attagtacat ttattaagcg ctagattctg tgcgttgttg
     4741 atttacagac aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg
     4801 gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa
     4861 atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga
     4921 accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg
     4981 tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac
     5041 gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg
     5101 actcgacgta aacacgttaa atagagcttg gacatattta acatcgggcg tgttagcttt
     5161 attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga
     5221 ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt
     5281 tgtagttgag ctttttggaa ttatttctga ttgcgggcgt ttttgggcgg gtttcaatct
     5341 aactgtgccc gattttaatt cagacaacac gttagaaagc gatggtgcag gcggtggtaa
     5401 catttcagac ggcaaatcta ctaatggcgg cggtggtgga gctgatgata aatctaccat
     5461 cggtggaggc gcaggcgggg ctggcggcgg aggcggaggc ggaggtggtg gcggtgatgc
     5521 agacggcggt ttaggctcaa atgtctcttt aggcaacaca gtcggcacct caactattgt
     5581 actggtttcg ggcgccgttt ttggtttgac cggtctgaga cgagtgcgat ttttttcgtt
     5641 tctaatagct tccaacaatt gttgtctgtc gtctaaaggt gcagcgggtt gaggttccgt
     5701 cggcattggt ggagcgggcg gcaattcaga catcgatggt ggtggtggtg gtggaggcgc
     5761 tggaatgtta ggcacgggag aaggtggtgg cggcggtgcc gccggtataa tttgttctgg
     5821 tttagtttgt tcgcgcacga ttgtgggcac cggcgcaggc gccgctggct gcacaacgga
     5881 aggtcgtctg cttcgaggca gcgcttgggg tggtggcaat tcaatattat aattggaata
     5941 caaatcgtaa aaatctgcta taagcattgt aatttcgcta tcgtttaccg tgccgatatt
     6001 taacaaccgc tcaatgtaag caattgtatt gtaaagagat tgtctcaagc tcggatcgat
     6061 cccgcacgcc gataacaagc cttttcattt ttactacagc attgtagtgg cgagacactt
     6121 cgctgtcgtc gaggtttaaa cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6181 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6241 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6301 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6361 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6421 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6481 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6541 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6601 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6661 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6721 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6781 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6841 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6901 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6961 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7021 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7081 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7141 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7201 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7261 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7321 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7381 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7441 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7501 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7561 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7621 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7681 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7741 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7801 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7861 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7921 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7981 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8041 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     8101 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8161 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8221 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8281 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8341 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8401 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     8461 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     8521 atttcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     8581 tcgctattac gcca
//
