LOCUS       sequence-52206-e        8948 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 8948)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8948)
  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            "Unpublished"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_2350
FEATURES             Location/Qualifiers
     source          1..8948
                     /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             4998..5015
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_recomb     5052..6409
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(5068..6409)
                     /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(6501..6518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6672..7260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6752..6771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7431..8222,8223..8291))
                     /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     8054..8073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8292..8396)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8423..8878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8510..8529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8720..8741
                     /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 aagaagcgtc acactctcga aaagatgact gactccgtga cctgtctgta ctgcaattcc
     4681 ttctccaagc aaagcaagca gaagaatttc ctgctcgtag ggaccgctga cggaaaactg
     4741 gcaatcttcg aggacaaaac cgtgaagttg aagggtgccg caccccttaa gatcctgaac
     4801 attggcaacg ttagcacccc tctgatgtgt ctatcagaat caactaactc caccgagcgc
     4861 aatgtgatgt ggggcgggtg cggcaccaag attttctcct tcagcaacga cttcaccatc
     4921 cagaagctga tcgagacccg taccagtcag ctcttcagtt acgccgcctt ctcctaaagc
     4981 ttgcggccgc actcgagcac caccaccacc accactaacc taggtagctg agcgcatgca
     5041 agctgatccg ggttattagt acatttatta agcgctagat tctgtgcgtt gttgatttac
     5101 agacaattgt tgtacgtatt ttaataattc attaaattta taatctttag ggtggtatgt
     5161 tagagcgaaa atcaaatgat tttcagcgtc tttatatctg aatttaaata ttaaatcctc
     5221 aatagatttg taaaataggt ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat
     5281 ggctggacta tctaatggat tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag
     5341 caatctagct ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt
     5401 caaaatatta tgcgcttttg tatttctttc atcactgtcg ttagtgtaca attgactcga
     5461 cgtaaacacg ttaaatagag cttggacata tttaacatcg ggcgtgttag ctttattagg
     5521 ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag acgattttgc
     5581 catagccaca cgacgcctat taattgtgtc ggctaacacg tccgcgatca aatttgtagt
     5641 tgagcttttt ggaattattt ctgattgcgg gcgtttttgg gcgggtttca atctaactgt
     5701 gcccgatttt aattcagaca acacgttaga aagcgatggt gcaggcggtg gtaacatttc
     5761 agacggcaaa tctactaatg gcggcggtgg tggagctgat gataaatcta ccatcggtgg
     5821 aggcgcaggc ggggctggcg gcggaggcgg aggcggaggt ggtggcggtg atgcagacgg
     5881 cggtttaggc tcaaatgtct ctttaggcaa cacagtcggc acctcaacta ttgtactggt
     5941 ttcgggcgcc gtttttggtt tgaccggtct gagacgagtg cgattttttt cgtttctaat
     6001 agcttccaac aattgttgtc tgtcgtctaa aggtgcagcg ggttgaggtt ccgtcggcat
     6061 tggtggagcg ggcggcaatt cagacatcga tggtggtggt ggtggtggag gcgctggaat
     6121 gttaggcacg ggagaaggtg gtggcggcgg tgccgccggt ataatttgtt ctggtttagt
     6181 ttgttcgcgc acgattgtgg gcaccggcgc aggcgccgct ggctgcacaa cggaaggtcg
     6241 tctgcttcga ggcagcgctt ggggtggtgg caattcaata ttataattgg aatacaaatc
     6301 gtaaaaatct gctataagca ttgtaatttc gctatcgttt accgtgccga tatttaacaa
     6361 ccgctcaatg taagcaattg tattgtaaag agattgtctc aagctcggat cgatcccgca
     6421 cgccgataac aagccttttc atttttacta cagcattgta gtggcgagac acttcgctgt
     6481 cgtcgaggtt taaacgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7081 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7561 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8401 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     8461 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     8521 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     8581 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     8641 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     8701 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     8761 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     8821 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc
     8881 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     8941 ttacgcca
//
