LOCUS       sequence-52222-e        8518 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 8518)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8518)
  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            "Unpublished"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: pEMB52 / Human_LRRK1_1_360
FEATURES             Location/Qualifiers
     source          1..8518
                     /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             4568..4585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_recomb     4622..5979
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4638..5979)
                     /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(6071..6088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6242..6830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6322..6341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7001..7792,7793..7861))
                     /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     7624..7643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7862..7966)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7993..8448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8080..8099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8290..8311
                     /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 ccggactgcc tgtcccgctg
     3481 caggagctgg ttcctgaact gttcatgacc gatttccccg ctcgcttgtt tctggagaat
     3541 agtaaattgg aacactcaga ggatgagggt agcgtgctgg ggcaaggtgg gtcagggacc
     3601 gtgatctatc gtgcgcgtta ccaaggacag cccgtcgctg tgaagcgctt tcatatcaag
     3661 aagttcaaaa atttcgcaaa tgtgcctgct gatacgatgt tacgtcatct tcgggctaca
     3721 gatgccatga aaaatttctc tgaatttcgg caagaagcct ctatgttaca tgccctgcaa
     3781 cacccgtgta tcgtggcctt gatcggcatt agcattcatc cgctctgctt cgccttagaa
     3841 ctggcaccgc tgtcttccct gaataccgta ttaagtgaaa acgcccgtga cagctcgttc
     3901 atcccactgg gtcatatgct tacccagaag atcgcatatc aaatagcgtc gggcctggca
     3961 tatctgcaca agaaaaacat tattttttgt gatttgaaat cggacaacat tttagtgtgg
     4021 tcgctagacg tgaaagaaca catcaacatt aaactcagcg actacggtat ttcgcgccag
     4081 agctttcacg aaggagcgtt gggcgtagaa ggtactccag gttatcaagc accggaaatt
     4141 cgcccaagaa ttgtatatga tgaaaaagtc gatatgttct cttacggtat ggtactgtac
     4201 gaactgctta gtggccagcg gccggcgctc ggccatcatc aactccagat tgccaaaaaa
     4261 ctcagcaaag ggattagacc ggtgcttggc caaccggagg aagttcagtt tcgacgtcta
     4321 caggcgctca tgatggagtg ttgggacact aaaccggaaa aacgcccact ggcgctgtcc
     4381 gttgtttccc agatgaaaga tccgacgttt gccacattta tgtacgagct gtgctgcggc
     4441 aaacagacgg cgtttttttc aagccagggc caggaatata ccgtcgtttt ctgggatggc
     4501 aaagaagaaa gtcgcaacta cacggtcgtt aacacggaaa aatgataagc ttgcggccgc
     4561 actcgagcac caccaccacc accactaacc taggtagctg agcgcatgca agctgatccg
     4621 ggttattagt acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt
     4681 tgtacgtatt ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa
     4741 atcaaatgat tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg
     4801 taaaataggt ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta
     4861 tctaatggat tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct
     4921 ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta
     4981 tgcgcttttg tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg
     5041 ttaaatagag cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg
     5101 tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca
     5161 cgacgcctat taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt
     5221 ggaattattt ctgattgcgg gcgtttttgg gcgggtttca atctaactgt gcccgatttt
     5281 aattcagaca acacgttaga aagcgatggt gcaggcggtg gtaacatttc agacggcaaa
     5341 tctactaatg gcggcggtgg tggagctgat gataaatcta ccatcggtgg aggcgcaggc
     5401 ggggctggcg gcggaggcgg aggcggaggt ggtggcggtg atgcagacgg cggtttaggc
     5461 tcaaatgtct ctttaggcaa cacagtcggc acctcaacta ttgtactggt ttcgggcgcc
     5521 gtttttggtt tgaccggtct gagacgagtg cgattttttt cgtttctaat agcttccaac
     5581 aattgttgtc tgtcgtctaa aggtgcagcg ggttgaggtt ccgtcggcat tggtggagcg
     5641 ggcggcaatt cagacatcga tggtggtggt ggtggtggag gcgctggaat gttaggcacg
     5701 ggagaaggtg gtggcggcgg tgccgccggt ataatttgtt ctggtttagt ttgttcgcgc
     5761 acgattgtgg gcaccggcgc aggcgccgct ggctgcacaa cggaaggtcg tctgcttcga
     5821 ggcagcgctt ggggtggtgg caattcaata ttataattgg aatacaaatc gtaaaaatct
     5881 gctataagca ttgtaatttc gctatcgttt accgtgccga tatttaacaa ccgctcaatg
     5941 taagcaattg tattgtaaag agattgtctc aagctcggat cgatcccgca cgccgataac
     6001 aagccttttc atttttacta cagcattgta gtggcgagac acttcgctgt cgtcgaggtt
     6061 taaacgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6121 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6181 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6241 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6301 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6361 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6421 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6481 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6541 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6601 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6661 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6721 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6781 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6841 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6901 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6961 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7021 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7081 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7141 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7201 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7261 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7321 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7381 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7441 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7501 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7561 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7621 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7681 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7741 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7801 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7861 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7921 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7981 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     8041 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     8101 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     8161 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     8221 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     8281 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     8341 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     8401 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc cattcgccat
     8461 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgcca
//
