LOCUS       sequence-324321-        5001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RAB35 in mammalian cells with a 2xmyc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5001)
  AUTHORS   Bonet-Ponce L, Beilina A, Williamson CD, Lindberg E, Kluss JH, 
            Saez-Atienzar S, Landeck N, Kumaran R, Mamais A, Bleck CKE, Li Y, 
            Cookson MR
  TITLE     LRRK2 mediates tubulation and vesicle sorting from lysosomes.
  JOURNAL   Sci Adv. 2020 Nov 11;6(46). pii: 6/46/eabb2454. doi: 
            10.1126/sciadv.abb2454. Print 2020 Nov.
  PUBMED    33177079
REFERENCE   2  (bases 1 to 5001)
  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: "2020-11-11"; volume: "6(46). pii: 6/46/eabb2454. doi"; pages:
            " 10.1126/sciadv.abb2454. Print 2020 Nov"
COMMENT     Sequence Label: 2xmyc-RAB35
FEATURES             Location/Qualifiers
     source          1..5001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             679..708
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             724..753
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    772..796
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1429..1453)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1502..1521)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1503..1521)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1795..1916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1832..1851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1886..1905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1923..2378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2060..2081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2272..2291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2405..2509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2511..2868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2511..2702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2534..2554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2719..2854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2903..3697
                     /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             2903..3697
                     /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(2957..2976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3567..3586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3885..3904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3929..3976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4305..4893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4794..4813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccatgga gcagaaactc atctctgaag aggatctgag cccgggcgga
      721 tccgagcaga aactcatctc tgaagaggat ctgctgcagg aattcgatat cacaagtttg
      781 tacaaaaaag caggctccga attcgccctt atggcccggg actacgacca cctcttcaag
      841 ctgctcatca tcggcgacag cggtgtgggc aagagcagtt tactgttgcg ttttgcagac
      901 aacactttct caggcagcta catcaccacg atcggagtgg atttcaagat ccggaccgtg
      961 gagatcaacg gggagaaggt gaagctgcag atctgggaca cagcggggca ggagcgcttc
     1021 cgcaccatca cctccacgta ttatcggggg acccacgggg tcattgtggt ttacgacgtc
     1081 accagtgccg agtcctttgt caacgtcaag cggtggcttc acgaaatcaa ccagaactgt
     1141 gatgatgtgt gccgaatatt agtgggtaat aagaatgacg accctgagcg gaaggtggtg
     1201 gagacggaag atgcctacaa attcgccggg cagatgggca tccagttgtt cgagaccagc
     1261 gccaaggaga atgtcaacgt ggaagagatg ttcaactgca tcacggagct ggtcctccga
     1321 gcaaagaaag acaacctggc aaaacagcag cagcaacaac agaacgatgt ggtgaagctc
     1381 acgaagaaca gtaaacgaaa gaaacgctgc tgctaaaggg cgaattcgac ccagctttct
     1441 tgtacaaagt ggtgatcggt accttaatta attaaggtac caggtaagtg tacccaattc
     1501 gccctatagt gagtcgtatt acaattcact cgatcgccct tcccaacagt tgcgcagcct
     1561 gaatggcgaa tggagatcca atttttaagt gtataatgtg ttaaactact gattctaatt
     1621 gtttgtgtat tttagattca cagtcccaag gctcatttca ggcccctcag tcctcacagt
     1681 ctgttcatga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1741 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1801 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1861 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     1921 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1981 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2041 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2101 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2161 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2221 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2281 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2341 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaatc ctgaggcgga
     2521 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2821 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     2881 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2941 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3001 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3061 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3121 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3181 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3241 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3301 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3361 cgatcaggat gatctggacg aagaacatca ggggctcgcg ccagccgaac tgttcgccag
     3421 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3481 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3541 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagaacttgg
     3601 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3661 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3721 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3841 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     3901 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     3961 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4021 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4081 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4141 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4441 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4981 gtggataacc gtattaccgc c
//
