LOCUS       sequence-414913-        8002 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 8002)
  TITLE     The Parkinson Disease-associated Leucine-rich Repeat Kinase 2 
            (LRRK2) Is a Dimer That Undergoes Intramolecular Autophosphorylation
  JOURNAL   J Biol Chem. 2008 Jun 13;283(24):16906-14.
  PUBMED    18397888
REFERENCE   2  (bases 1 to 8002)
  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: "J Biol 
            Chem."; date: "2008-06-13"; volume: "283(24)"; pages: "16906-14"
COMMENT     Sequence Label: 2XMyc-LRRK2-RCKW
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        92..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        645..663
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      695..704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             704..733
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             749..778
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    797..821
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4455..4479)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4528..4547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4529..4547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4821..4942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4858..4877)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4912..4931
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4949..5404)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5086..5107)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5298..5317
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5431..5535
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5537..5894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5537..5728
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5560..5580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5745..5880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5929..6723
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5929..6723
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5983..6002)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6593..6612
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6911..6930)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6955..7002
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7331..7919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7820..7839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      601 gtttagtgaa ccgtcagatc cgctagcgat tacgccaagc tcgaaattaa ccctcactaa
      661 agggaacaaa agctggagct ccaccgcggt ggcggccgcc atggagcaga aactcatctc
      721 tgaagaggat ctgagcccgg gcggatccga gcagaaactc atctctgaag aggatctgct
      781 gcaggaattc gatatcacaa gtttgtacaa aaaagcaggc tccgaattcg cccttatgaa
      841 ggctgtgcct tataaccgaa tgaaacttat gattgtggga aatactggga gtggtaaaac
      901 caccttattg cagcaattaa tgaaaaccaa gaaatcagat cttggaatgc aaagtgccac
      961 agttggcata gatgtgaaag actggcctat ccaaataaga gacaaaagaa agagagatct
     1021 cgtcctaaat gtgtgggatt ttgcaggtcg tgaggaattc tatagtactc atccccattt
     1081 tatgacgcag cgagcattgt accttgctgt ctatgacctc agcaagggac aggctgaagt
     1141 tgatgccatg aagccttggc tcttcaatat aaaggctcgc gcttcttctt cccctgtgat
     1201 tctcgttggc acacatttgg atgtttctga tgagaagcaa cgcaaagcct gcatgagtaa
     1261 aatcaccaag gaactcctga ataagcgagg gttccctgcc atacgagatt accactttgt
     1321 gaatgccacc gaggaatctg atgctttggc aaaacttcgg aaaaccatca taaacgagag
     1381 ccttaatttc aagatccgag atcagcttgt tgttggacag ctgattccag actgctatgt
     1441 agaacttgaa aaaatcattt tatcggagcg taaaaatgtg ccaattgaat ttcccgtaat
     1501 tgaccggaaa cgattattac aactagtgag agaaaatcag ctgcagttag atgaaaatga
     1561 gcttcctcac gcagttcact ttctaaatga atcaggagtc cttcttcatt ttcaagaccc
     1621 agcactgcag ttaagtgact tgtactttgt ggaacccaag tggctttgta aaatcatggc
     1681 acagattttg acagtgaaag tggaaggttg tccaaaacac cctaagggaa ttatttcgcg
     1741 tagagatgtg gaaaaatttc tttcaaaaaa aaggaaattt ccaaagaact acatgtcaca
     1801 gtattttaag ctcctagaaa aattccagat tgctttgcca ataggagaag aatatttgct
     1861 ggttccaagc agtttgtctg accacaggcc tgtgatagag cttccccatt gtgagaactc
     1921 tgaaattatc atccgactat atgaaatgcc ttattttcca atgggatttt ggtcaagatt
     1981 aatcaatcga ttacttgaga tttcacctta catgctttca gggagagaac gagcacttcg
     2041 cccaaacaga atgtattggc gacaaggcat ttacttaaat tggtctcctg aagcttattg
     2101 tctggtagga tctgaagtct tagacaatca tccagagagt ttcttaaaaa ttacagttcc
     2161 ttcttgtaga aaaggctgta ttcttttggg ccaagttgtg gaccacattg attctctcat
     2221 ggaagaatgg tttcctgggt tgctggagat tgatatttgt ggtgaaggag aaactctgtt
     2281 gaagaaatgg gcattatata gttttaatga tggcgaagaa catcaaaaaa tcttacttga
     2341 tgacttgatg aagaaagcag aggaaggaga tctcttagta aatccagatc aaccaaggct
     2401 caccattcca atatctcaga ttgcccctga cttgattttg gctgacctgc ctagaaatat
     2461 tatgttgaat aatgatgagt tggaatttga acaagctcca gagtttctcc taggtgatgg
     2521 cagttttgga tcagtttacc gagcagccta tgaaggagaa gaagtggctg tgaagatttt
     2581 taataaacat acatcactca ggctgttaag acaagagctt gtggtgcttt gccacctcca
     2641 ccaccccagt ttgatatctt tgctggcagc tgggattcgt ccccggatgt tggtgatgga
     2701 gttagcctcc aagggttcct tggatcgcct gcttcagcag gacaaagcca gcctcactag
     2761 aaccctacag cacaggatcg cactccacgt agctgatggt ttgagatacc tccactcagc
     2821 catgattata taccgagacc tgaaacccca caatgtgctg cttttcacac tgtatcccaa
     2881 tgctgccatc atcgcaaaga ttgctgacta cggcattgct cagtactgct gtagaatggg
     2941 gataaaaaca tcagagggca caccagggtt tcgtgcacct gaagttgcca gaggaaatgt
     3001 catttataac caacaggctg atgtttattc atttggttta ctactctatg acattttgac
     3061 aactggaggt agaatagtag agggtttgaa gtttccaaat gagtttgatg aattagaaat
     3121 acaaggaaaa ttacctgatc cagttaaaga atatggttgt gccccatggc ctatggttga
     3181 gaaattaatt aaacagtgtt tgaaagaaaa tcctcaagaa aggcctactt ctgcccaggt
     3241 ctttgacatt ttgaattcag ctgaattagt ctgtctgacg agacgcattt tattacctaa
     3301 aaacgtaatt gttgaatgca tggttgctac acatcacaac agcaggaatg caagcatttg
     3361 gctgggctgt gggcacaccg acagaggaca gctctcattt cttgacttaa atactgaagg
     3421 atacacttct gaggaagttg ctgatagtag aatattgtgc ttagccttgg tgcatcttcc
     3481 tgttgaaaag gaaagctgga ttgtgtctgg gacacagtct ggtactctcc tggtcatcaa
     3541 taccgaagat gggaaaaaga gacataccct agaaaagatg actgattctg tcacttgttt
     3601 gtattgcaat tccttttcca agcaaagcaa acaaaaaaat tttcttttgg ttggaaccgc
     3661 tgatggcaag ttagcaattt ttgaagataa gactgttaag cttaaaggag ctgctccttt
     3721 gaagatacta aatataggaa atgtcagtac tccattgatg tgtttgagtg aatccacaaa
     3781 ttcaacggaa agaaatgtaa tgtggggagg atgtggcaca aagattttct ccttttctaa
     3841 tgatttcacc attcagaaac tcattgagac aagaacaagc caactgtttt cttatgcagc
     3901 tttcagtgat tccaacatca taacagtggt ggtagacact gctctctata ttgctaagca
     3961 aaatagccct gttgtggaag tgtgggataa gaaaactgaa aaactctgtg gactaataga
     4021 ctgcgtgcac tttttaaggg aggtaatggt aaaagaaaac aaggaatcaa aacacaaaat
     4081 gtcttattct gggagagtga aaaccctctg ccttcagaag aacactgctc tttggatagg
     4141 aactggagga ggccatattt tactcctgga tctttcaact cgtcgactta tacgtgtaat
     4201 ttacaacttt tgtaattcgg tcagagtcat gatgacagca cagctaggaa gccttaaaaa
     4261 tgtcatgctg gtattgggct acaaccggaa aaatactgaa ggtacacaaa agcagaaaga
     4321 gatacaatct tgcttgaccg tttgggacat caatcttcca catgaagtgc aaaatttaga
     4381 aaaacacatt gaagtgagaa aagaattagc tgaaaaaatg agacgaacat ctgttgagta
     4441 aaagggcgaa ttcgacccag ctttcttgta caaagtggtg atcggtacct taattaatta
     4501 aggtaccagg taagtgtacc caattcgccc tatagtgagt cgtattacaa ttcactcgat
     4561 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga gatccaattt ttaagtgtat
     4621 aatgtgttaa actactgatt ctaattgttt gtgtatttta gattcacagt cccaaggctc
     4681 atttcaggcc cctcagtcct cacagtctgt tcatgatcat aatcagccat accacatttg
     4741 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4801 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4861 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4921 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     4981 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5041 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5101 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5161 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5221 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5281 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5341 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5401 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5521 tgaaaaagga agaatcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     5581 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5641 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5701 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5761 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5821 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5881 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5941 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6001 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6061 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     6121 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6181 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6241 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6301 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6361 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga acatcagggg
     6421 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     6481 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6541 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6601 acccgtgata ttgctgaaga acttggcggc gaatgggctg accgcttcct cgtgctttac
     6661 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6721 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6781 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6841 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     6901 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     6961 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     7021 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     7081 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     7141 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     7201 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7921 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7981 tctttcctgc gttatcccct ga
//
