LOCUS       sequence-398445-        6639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression in cultured cells to visualize RILPL1 protein 
            interactions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6639)
  AUTHORS   Pal P, Taylor M, Lam PY, Tonelli F, Hecht CA, Lis P, Nirujogi RS, 
            Phung TK, Yeshaw WM, Jaimon E, Fasimoye R, Dickie EA, Wightman M, 
            Macartney T, Pfeffer SR, Alessi DR
  TITLE     Parkinson's VPS35[D620N] mutation induces LRRK2-mediated lysosomal 
            association of RILPL1 and TMEM55B.
  JOURNAL   Sci Adv. 2023 Dec 15;9(50):eadj1205. doi: 10.1126/sciadv.adj1205. 
            Epub 2023 Dec 13.
  PUBMED    38091401
REFERENCE   2  (bases 1 to 6639)
  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: "Sci Adv."; 
            date: "2023-12-15"; volume: "9(50):eadj1205. doi"; pages: " 
            10.1126/sciadv.adj1205. Epub 2023 Dec 13"
COMMENT     Sequence Label: Myc-RILPL1
FEATURES             Location/Qualifiers
     source          1..6639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(103..207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(282..301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        473..852
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        853..1056
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1007..1027
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1101..1120
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1101..1119
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1142..1171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2451..2469)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2452..2469)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2489..2506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2495..2719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2765..3193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2852..2871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3062..3083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3202..3222)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3367..3501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3428..3447
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3582..4376
                     /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(3636..3655)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4246..4265
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4550..4671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4587..4606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4641..4660
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4720..4736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4720..4736)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4733..4755)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4744..4760
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4768..4774,4775..4792,4793..4798))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4813..4834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4951..4968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5122..5710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5202..5221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5881..6639,1..33,34..102))
                     /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     6504..6523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
       61 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
      121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
      241 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
      301 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      361 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      421 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      481 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      541 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      601 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      661 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      721 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      781 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      841 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      901 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      961 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1021 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     1081 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccat
     1141 ggaacaaaaa ctcatctcag aagaggatct gaatttatca attaccggtg gatcccccgg
     1201 ggaattcatg gaggaggagc gggggtcggc gctggcggcc gagtcggcgc tggagaagaa
     1261 cgtggccgag ctgaccgtca tggacgtgta cgacatcgcg tcgcttgtgg gccacgagtt
     1321 cgagcgggtc attgaccagc acggctgcga ggccatcgcg cgcctcatgc ccaaggtcgt
     1381 gcgcgtcctg gagatcctgg aggtgctggt cagccgccac cacgtcgcgc ccgagctgga
     1441 cgagctgcgc ctggagctgg accgcctgcg cctggagagg atggaccgca tcgagaagga
     1501 gcgcaagcac cagaaggagc tggagctggt ggaggatgtg tggcgagggg aggcgcagga
     1561 cctcctctcc cagatcgccc agctgcagga ggagaacaag cagctcatga ccaacctctc
     1621 ccacaaggat gtcaacttct cagaggagga gttccagaag catgaaggca tgtcagagcg
     1681 ggagcgacag gtgatgaaga agctgaagga ggtggtggac aaacaacgcg acgagatccg
     1741 cgccaaggac agggagctgg gcctgaaaaa tgaggacgtt gaggctttac agcagcagca
     1801 gacacggctg atgaagatca accatgacct tcggcaccgg gtcacggtgg tggaggccca
     1861 ggggaaagcc ctgatcgaac agaaggtgga gctggaggca gacctgcaga ccaaggagca
     1921 ggagatgggc agcctgcgag cagagctggg gaagttgcga gagaggctgc agggggagca
     1981 cagccagaat ggggaggagg agcctgagac ggagccggtg ggagaggaga gcatctccga
     2041 cgcagagaag gtggccatgg atctcaagga ccccaaccgc ccccggttca ccctgcagga
     2101 gctgcgggac gtgctgcacg agaggaacga gctcaagtcc aaggtgttct tgctgcagga
     2161 ggagctggct tactataaga gtgaagaaat ggaagaggaa aaccgaatac cccaaccccc
     2221 acccatcgcc cacccgagga cgtcccccca gccggagtcg ggcatcaagc gactgtttag
     2281 cttcttctcc cgagataaga agcgcctggc caacacacag agaaacgtgc acatccagga
     2341 gtcctttgga cagtgggcaa acacccaccg cgatgacggt tacacagagc aaggacagga
     2401 agccctgcag catctggcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     2461 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2521 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2581 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2641 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2701 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2761 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2821 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2881 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2941 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3001 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3061 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3121 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3181 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3301 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3361 ccatagtccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3421 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3481 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3541 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3601 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3661 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3721 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3781 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3841 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3901 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3961 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4021 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4081 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4141 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4201 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4261 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4321 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4381 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4441 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4501 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4561 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4621 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4681 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4741 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4801 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4861 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4921 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4981 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5041 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5101 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5161 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5221 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5281 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5341 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5401 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5461 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5521 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5581 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5641 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5701 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5761 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5821 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5881 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5941 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6001 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6061 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6121 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6181 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6241 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6301 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6361 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6421 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6481 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6541 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6601 cagttcgatg taacccactc gtgcacccaa ctgatcttc
//
