LOCUS       sequence-52228-e        8476 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 8476)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8476)
  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            "Unpublished"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: pEMB44 / Mouse_LRRK2_1_342
FEATURES             Location/Qualifiers
     source          1..8476
                     /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             3433..3456
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3457..3474
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4526..4543
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_recomb     4580..5937
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4596..5937)
                     /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(6029..6046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6200..6788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6280..6299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6959..7750,7751..7819))
                     /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     7582..7601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7820..7924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7951..8406
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8038..8057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8248..8269
                     /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 aggatccagg
     3421 cctgaattca tgcaccacca ccatcaccat catcatttag ttcctcgcgg ctcaggcctg
     3481 acaatcccta tctctcagat agcaccggat ctaatcttgg ccgatttgcc gcgaaacatc
     3541 atgcttaata acgacgaact cgagttcgag gaagcgccgg aatttttgtt gggagatggc
     3601 tccttcggca gtgtctaccg tgcggcttat gagggtgagg aagttgctgt taaaatcttt
     3661 aataaacaca cctcgcttag actattgcga caagagctgg tggttctttg ccatctgcat
     3721 catccaagcc tcataagttt gttagcagcc ggtatccggc ctcgtatgtt agtgatggag
     3781 ttagcctcta aagggtccct cgaccgcctt ctccagcagg acaaagcaag tcttactcgt
     3841 accctccagc atcgtatcgc tctgcatgtg gctgacggct tacgctacct gcatagcgcc
     3901 atgattattt atcgcgatct gaaaccacat aacgtcttac tgtttactct gtaccccaat
     3961 gccgcaatca ttgcgaaaat cgccgattat ggcattgcac aatattgctg ccgtatgggc
     4021 attaagacca gcgagggtac accaggtttt cgcgcgcccg aagtggcacg tgggaacgtt
     4081 atctataatc agcaggcgga tgtatactcg tttgggttac tgctgcacga catctggaca
     4141 accggtagcc gcattatgga agggctgcgt ttcccgaacg aatttgatga actggcgatt
     4201 cagggtaaac tgccggatcc tgttaaggaa tacggctgcg caccgtggcc gatggtagaa
     4261 aaactgatta cgaaatgtct gaaagaaaac ccgcaagaac gcccgacgtc agcgcaggtg
     4321 ttcgatattc tgaatagcgc ggaactgatt tgtctgatgc gccacattct gattccgaaa
     4381 aacattattg tagaatgtat ggtcgcgacg aacctgaata gtaagtccgc gacgctgtgg
     4441 ctgggttgtg gaaataccga aaagggccaa ctgtcgctgt tcgatcttaa taccgaacgc
     4501 tgataagctt gcggccgcac tcgagcacca ccaccaccac cactaaccta ggtagctgag
     4561 cgcatgcaag ctgatccggg ttattagtac atttattaag cgctagattc tgtgcgttgt
     4621 tgatttacag acaattgttg tacgtatttt aataattcat taaatttata atctttaggg
     4681 tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt
     4741 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc
     4801 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct
     4861 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg
     4921 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat
     4981 tgactcgacg taaacacgtt aaatagagct tggacatatt taacatcggg cgtgttagct
     5041 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
     5101 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
     5161 tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc gggtttcaat
     5221 ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc aggcggtggt
     5281 aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga taaatctacc
     5341 atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg tggcggtgat
     5401 gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac ctcaactatt
     5461 gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg atttttttcg
     5521 tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg ttgaggttcc
     5581 gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg tggtggaggc
     5641 gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat aatttgttct
     5701 ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg ctgcacaacg
     5761 gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt ataattggaa
     5821 tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac cgtgccgata
     5881 tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa gctcggatcg
     5941 atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt ggcgagacac
     6001 ttcgctgtcg tcgaggttta aacgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6061 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6601 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7921 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     7981 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     8041 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8101 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8161 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8221 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8281 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     8341 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     8401 caatttccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     8461 cttcgctatt acgcca
//
