LOCUS       sequence-425850-        5489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human LRRK2 fragment R2142 to E2527. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5489)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 5489)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pBiT3.1-C_LRRK2:2142-2527
FEATURES             Location/Qualifiers
     source          1..5489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2552..2673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2563..2582)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2788..2821)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2831..2878
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2910..3704
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2964..2983)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3574..3593
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3862..4450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3942..3961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4576..4624
                     /note="synthetic polyadenylation signal"
     CDS             complement(4648..5046)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5043..5052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5077..5434)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5091..5226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5243..5434
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5391..5411
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgagacg cattttatta
     1261 cctaaaaacg taattgttga atgcatggtt gctacacatc acaacagcag gaatgcaagc
     1321 atttggctgg gctgtgggca caccgacaga ggacagctct catttcttga cttaaatact
     1381 gaaggataca cttctgagga agttgctgat agtagaatat tgtgcttagc cttggtgcat
     1441 cttcctgttg aaaaggaaag ctggattgtg tctgggacac agtctggtac tctcctggtc
     1501 atcaataccg aagatgggaa aaagagacat accctagaaa agatgactga ttctgtcact
     1561 tgtttgtatt gcaattcctt ttccaagcaa agcaaacaaa aaaattttct tttggttgga
     1621 accgctgatg gcaagttagc aatttttgaa gataagactg ttaagcttaa aggagctgct
     1681 cctttgaaga tactaaatat aggaaatgtc agtactccat tgatgtgttt gagtgaatcc
     1741 acaaattcaa cggaaagaaa tgtaatgtgg ggaggatgtg gcacaaagat tttctccttt
     1801 tctaatgatt tcaccattca gaaactcatt gagacaagaa caagccaact gttttcttat
     1861 gcagctttca gtgattccaa catcataaca gtggtggtag acactgctct ctatattgct
     1921 aagcaaaata gccctgttgt ggaagtgtgg gataagaaaa ctgaaaaact ctgtggacta
     1981 atagactgcg tgcacttttt aagggaggta acggtaaaag aaaacaagga atcaaaacac
     2041 aaaatgtctt attctgggag agtgaaaacc ctctgccttc agaagaacac tgctctttgg
     2101 ataggaactg gaggaggcca tattttactc ctggatcttt caactcgtcg acttatacgt
     2161 gtaatttaca acttttgtaa ttcggtcaga gtcatgatga cagcacagct aggaagcctt
     2221 aaaaatgtca tgctggtatt gggctacaac cggaaaaata ctgaaggtac acaaaagcag
     2281 aaagagatac aatcttgctt gaccgtttgg gacatcaatc ttccacatga agtgcaaaat
     2341 ttagaaaaac acattgaagt gagaaaagaa ttagctgaaa aaatgagacg aacatctgtt
     2401 gagggctcga gcggtgggag ctccggtgtg agcggctggc ggctgttcaa gaagattagc
     2461 taagtttaaa cggccgcgac tctagactgc aggcatgcaa gctgatttgg ctgctgccac
     2521 cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2581 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2641 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2701 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     2761 ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta tacgaagtta
     2821 tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     2881 ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt
     2941 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3001 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3061 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3121 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3181 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3241 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3301 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3361 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3421 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3481 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3541 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3601 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3661 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa atgaccgacc
     3721 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3781 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3841 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3901 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3961 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4021 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4081 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4201 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4261 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     4321 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4381 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4441 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4501 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4561 atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     4621 tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag agggcagcaa
     4681 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
     4741 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
     4801 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
     4861 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
     4921 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
     4981 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
     5041 ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     5101 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     5161 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     5221 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     5281 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     5341 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     5401 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     5461 agaggttatt tcaggccatg gtgctgcgc
//
