LOCUS       Exported                8464 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEMB44 / Zebrafish_LRRK2_1_338
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8464)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8464)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8464
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             3446..3469
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3470..3487
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4514..4531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_recomb     4568..5925
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4584..5925)
                     /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(6017..6034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6188..6776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6268..6287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6947..7807)
                     /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     7570..7589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7808..7912)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7939..8394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8026..8045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8236..8257
                     /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 cctgaattcg agctccaagc ttatgcacca ccaccatcac catcatcatt tagttcctcg
     3481 cggctcagga tgcacgttac ctatatcgca gatttcaccg gacctggtgc tttctgacca
     3541 gccttcctcg acaatcctgg atccagagca gttagaaatg gaactaaccg ctgagtatat
     3601 gctcggcgat ggaggcttcg gcagtgttta taaagccgtg tacaaaaacg aggaggtggc
     3661 ggtcaagatc tttaacaaac acgcatcggc tttgtatgtg caccgcttag tccgccagga
     3721 gctagccgta ttagggcgtc tctgtcatcc ctcgctggtc ggcctgttag cggccggatg
     3781 caatccccat atcctcgtaa tggagctggc accgtacggc agtctggact cgctcttcga
     3841 acgagaaaac ggttccttgt caagaaaatt gcaacaccgt attgcgttgc acgtcgccga
     3901 cgggcttaaa tacttgcata gttcaatgat catctaccgg gacctgaaac cgcataacgt
     3961 actgcttttt aatcttaaga ccgatgctga gattgttgca aaaattacgg attacggcat
     4021 cgcacagtat tgttgtagta tgggcgtccg gtcatctgaa ggaaccccgg gtttccgtgc
     4081 gcctgaagtg gcccgcggga acgtgattta taatgtgcag gcagatgttt atagcttcgg
     4141 tctgctgctg tatgacctgc tgacctatgg tgaacgcatt tccgatggca tgaaatttcc
     4201 gtctgaattc gatgaagtag ccgtccaggg caaactgccg gatccagtga aagattacgg
     4261 ctgctccccg tggccggaaa ttgaatctct gatgcgtgaa tgcatgcgcg aaaatccaca
     4321 agatcgcccg actagcgcgc aagtgtttga tcgtctgaac agcgcggaaa tgctgtgcct
     4381 gactcgtgaa ctgaatgtgg ttggttttcc aggcgaatgt tttgttgtta gcaacagcgg
     4441 tggcgcggcg aacgggggta agaatccgca tgtttggatt ggtggtggta gcagcagcca
     4501 atgataactc gagcaccacc accaccacca ctaacctagg tagctgagcg catgcaagct
     4561 gatccgggtt attagtacat ttattaagcg ctagattctg tgcgttgttg atttacagac
     4621 aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg gtatgttaga
     4681 gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa atcctcaata
     4741 gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga accgatggct
     4801 ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat
     4861 ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa
     4921 atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg actcgacgta
     4981 aacacgttaa atagagcttg gacatattta acatcgggcg tgttagcttt attaggccga
     5041 ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga ttttgccata
     5101 gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag
     5161 ctttttggaa ttatttctga ttgcgggcgt ttttgggcgg gtttcaatct aactgtgccc
     5221 gattttaatt cagacaacac gttagaaagc gatggtgcag gcggtggtaa catttcagac
     5281 ggcaaatcta ctaatggcgg cggtggtgga gctgatgata aatctaccat cggtggaggc
     5341 gcaggcgggg ctggcggcgg aggcggaggc ggaggtggtg gcggtgatgc agacggcggt
     5401 ttaggctcaa atgtctcttt aggcaacaca gtcggcacct caactattgt actggtttcg
     5461 ggcgccgttt ttggtttgac cggtctgaga cgagtgcgat ttttttcgtt tctaatagct
     5521 tccaacaatt gttgtctgtc gtctaaaggt gcagcgggtt gaggttccgt cggcattggt
     5581 ggagcgggcg gcaattcaga catcgatggt ggtggtggtg gtggaggcgc tggaatgtta
     5641 ggcacgggag aaggtggtgg cggcggtgcc gccggtataa tttgttctgg tttagtttgt
     5701 tcgcgcacga ttgtgggcac cggcgcaggc gccgctggct gcacaacgga aggtcgtctg
     5761 cttcgaggca gcgcttgggg tggtggcaat tcaatattat aattggaata caaatcgtaa
     5821 aaatctgcta taagcattgt aatttcgcta tcgtttaccg tgccgatatt taacaaccgc
     5881 tcaatgtaag caattgtatt gtaaagagat tgtctcaagc tcggatcgat cccgcacgcc
     5941 gataacaagc cttttcattt ttactacagc attgtagtgg cgagacactt cgctgtcgtc
     6001 gaggtttaaa cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6061 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6601 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6901 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6961 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7021 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7081 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7141 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7201 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7261 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7321 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7381 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7441 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7501 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7561 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7621 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7681 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7741 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7801 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7861 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7921 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     7981 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     8041 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     8101 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     8161 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     8221 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     8281 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     8341 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcccatt
     8401 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     8461 gcca
//
