LOCUS       sequence-52068-e        8653 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 8653)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8653)
  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            "Unpublished"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: pBACgus_Sumostar/LRRK2 1867-2161
FEATURES             Location/Qualifiers
     source          1..8653
                     /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             3424..3441
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3454..3741
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLTFLYDGIEIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             3748..3771
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3772..3792
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4703..4720
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_recomb     4757..6114
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4773..6114)
                     /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(6206..6223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6377..6965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6457..6476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7136..7927,7928..7996))
                     /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     7759..7778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7997..8101)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8128..8583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8215..8234)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8425..8446
                     /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 ggtcatcacc atcatcatca cgggtccctg caggactcag aagtcaatca agaagctaag
     3481 ccagaggtca agccagaagt caagcctgag actcacatca atttaaaggt gtccgatgga
     3541 tcttcagaga tcttcttcaa gatcaaaaag accactcctt taagaaggct gatggaagcg
     3601 ttcgctaaaa gacagggtaa ggaaatggac tccttaacgt tcttgtacga cggtattgaa
     3661 attcaagctg atcagacccc tgaagatttg gacatggagg ataacgatat tattgaggct
     3721 cacagagaac agattggagg atccatgcac caccatcacc accatcacca cgagaacctt
     3781 tatttccaag gaaacatcat gctcaacaac gatgagctcg aatttgaaca agctccagaa
     3841 ttcctgctcg gcgacggctc tttcggctcc gtataccgtg ctgcttatga aggcgaagaa
     3901 gtcgccgtaa agatcttcaa caagcacacc tcgctccgtc tgctccgtca ggaactggtc
     3961 gttctgtgtc atctgcacca tccgagcctc atctccctgc tggccgctgg catccgtcct
     4021 cgtatgctgg tgatggagct ggcatccaag ggctctctcg accgcctgct gcagcaggac
     4081 aaggcttctc tgacccgtac tctgcagcat cgtatcgctc tgcatgtagc tgacggcctg
     4141 cgttatcttc attctgcaat gatcatttat cgcgacctga aacctcataa tgtactgctg
     4201 tttactctgt acccgaacgc tgccatcatc gctaagattg cagactatgg tattgcgcag
     4261 tactgctgcc gcatgggcat taaaacaagc gaaggtacgc cgggtttccg tgctccagaa
     4321 gtggctcgtg gcaacgttat ttacaaccag caagctgacg tttactcgtt cggtctgctc
     4381 ctgtacgata tcctgactac cggcggccgt atcgtagaag gtctgaagtt ccctaacgaa
     4441 ttcgacgaac tggaaatcca gggcaagctg ccagacccgg ttaaagaata cggttgcgct
     4501 ccgtggccga tggtagagaa gctcatcaaa cagtgcctta aagaaaaccc tcaggaacgt
     4561 ccaacctctg ctcaagtgtt cgacatcctt aacagcgctg aactcgtctg tctgactcgc
     4621 cgtattctcc tgcctaagaa tgttatcgtg gaatgtatgg ttgctactca ccataactga
     4681 taagcttgcg gccgcactcg agcaccacca ccaccaccac taacctaggt agctgagcgc
     4741 atgcaagctg atccgggtta ttagtacatt tattaagcgc tagattctgt gcgttgttga
     4801 tttacagaca attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg
     4861 tatgttagag cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa
     4921 tcctcaatag atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa
     4981 ccgatggctg gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt
     5041 agcagcaatc tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg
     5101 tcgttcaaaa tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga
     5161 ctcgacgtaa acacgttaaa tagagcttgg acatatttaa catcgggcgt gttagcttta
     5221 ttaggccgat tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat
     5281 tttgccatag ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt
     5341 gtagttgagc tttttggaat tatttctgat tgcgggcgtt tttgggcggg tttcaatcta
     5401 actgtgcccg attttaattc agacaacacg ttagaaagcg atggtgcagg cggtggtaac
     5461 atttcagacg gcaaatctac taatggcggc ggtggtggag ctgatgataa atctaccatc
     5521 ggtggaggcg caggcggggc tggcggcgga ggcggaggcg gaggtggtgg cggtgatgca
     5581 gacggcggtt taggctcaaa tgtctcttta ggcaacacag tcggcacctc aactattgta
     5641 ctggtttcgg gcgccgtttt tggtttgacc ggtctgagac gagtgcgatt tttttcgttt
     5701 ctaatagctt ccaacaattg ttgtctgtcg tctaaaggtg cagcgggttg aggttccgtc
     5761 ggcattggtg gagcgggcgg caattcagac atcgatggtg gtggtggtgg tggaggcgct
     5821 ggaatgttag gcacgggaga aggtggtggc ggcggtgccg ccggtataat ttgttctggt
     5881 ttagtttgtt cgcgcacgat tgtgggcacc ggcgcaggcg ccgctggctg cacaacggaa
     5941 ggtcgtctgc ttcgaggcag cgcttggggt ggtggcaatt caatattata attggaatac
     6001 aaatcgtaaa aatctgctat aagcattgta atttcgctat cgtttaccgt gccgatattt
     6061 aacaaccgct caatgtaagc aattgtattg taaagagatt gtctcaagct cggatcgatc
     6121 ccgcacgccg ataacaagcc ttttcatttt tactacagca ttgtagtggc gagacacttc
     6181 gctgtcgtcg aggtttaaac gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6241 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6301 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6361 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6421 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6481 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6541 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6601 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6661 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6721 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6781 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6841 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6901 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6961 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7021 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7081 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7141 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7201 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7261 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7321 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7381 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7441 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7501 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7561 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7621 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7681 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7741 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7801 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7861 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7921 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7981 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8041 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8101 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     8161 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     8221 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     8281 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     8341 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     8401 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     8461 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     8521 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     8581 tttcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     8641 cgctattacg cca
//
