LOCUS       sequence-489405-        5608 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Express RILPL1 in mammalian cells with 2xmyc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  AUTHORS   Bonet-Ponce L, Tegicho T, Fernandez-Martinez N, Rozenberg IA, 
            Ashriem M, Beilina A, Kluss JH, Li Y, Cookson MR
  TITLE     JIP4 and RILPL1 utilize opposing motor force to dynamically regulate
            lysosomal tubulation.
  JOURNAL   J Cell Biol. 2025 Nov 3;224(11):e202404018. doi: 
            10.1083/jcb.202404018. Epub 2025 Sep 24.
  PUBMED    40990465
REFERENCE   2  (bases 1 to 5608)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2025-11-3"; volume: "224(11):e202404018. doi"; pages:
            " 10.1083/jcb.202404018. Epub 2025 Sep 24"
COMMENT     Sequence Label: 2xmyc-RILPL1
FEATURES             Location/Qualifiers
     source          1..5608
                     /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(2036..2060)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2109..2128)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2110..2128)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2402..2523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2439..2458)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2493..2512
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2530..2985)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2667..2688)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2879..2898
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3012..3116
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3118..3475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3118..3309
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3141..3161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3326..3461
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3510..4304
                     /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             3510..4304
                     /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(3564..3583)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4174..4193
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4492..4511)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4536..4583
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4912..5500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5401..5420
                     /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 atggaggagg agcgggggtc ggcgctggcg
      841 gccgagtcgg cgctggagaa gaacgtggcc gagctgaccg tcatggacgt gtacgacatc
      901 gcgtcgcttg tgggccacga gttcgagcgg gtcattgacc agcacggctg cgaggccatc
      961 gcgcgcctca tgcccaaggt cgtgcgcgtc ctggagatcc tggaggtgct ggtcagccgc
     1021 caccacgtcg cgcccgagct ggacgagctg cgcctggagc tggaccgcct gcgcctggag
     1081 aggatggacc gcatcgagaa ggagcgcaag caccagaagg agctggagct ggtggaggat
     1141 gtgtggcgag gggaggcgca ggacctcctc tcccagatcg cccagctgca ggaggagaac
     1201 aagcagctca tgaccaacct ctcccacaag gatgtcaact tctcagagga ggagttccag
     1261 aagcatgaag gcatgtcaga gcgggagcga caggtgatga agaagctgaa ggaggtggtg
     1321 gacaaacaac gcgacgagat ccgcgccaag gacagggagc tgggcctgaa aaatgaggac
     1381 gttgaggctt tacagcagca gcagacacgg ctgatgaaga tcaaccatga ccttcggcac
     1441 cgggtcacgg tggtggaggc ccaggggaaa gccctgatcg aacagaaggt ggagctggag
     1501 gcagacctgc agaccaagga gcaggagatg ggcagcctgc gagcagagct ggggaagttg
     1561 cgagagaggc tgcaggggga gcacagccag aatggggagg aggagcctga gacggagccg
     1621 gtgggagagg agagcatctc cgacgcagag aaggtggcca tggatctcaa ggaccccaac
     1681 cgcccccggt tcaccctgca ggagctgcgg gacgtgctgc acgagaggaa cgagctcaag
     1741 tccaaggtgt tcttgctgca ggaggagctg gcttactata agagtgaaga aatggaagag
     1801 gaaaaccgaa taccccaacc cccacccatc gcccacccga ggacgtcccc ccagccggag
     1861 tcgggcatca agcgactgtt tagcttcttc tcccgagata agaagcgcct ggccaacaca
     1921 cagagaaacg tgcacatcca ggagtccttt ggacagtggg caaacaccca ccgcgatgac
     1981 ggttacacag agcaaggaca ggaagccctg cagcatctgt gaaagggcga attcgaccca
     2041 gctttcttgt acaaagtggt gatcggtacc ttaattaatt aaggtaccag gtaagtgtac
     2101 ccaattcgcc ctatagtgag tcgtattaca attcactcga tcgcccttcc caacagttgc
     2161 gcagcctgaa tggcgaatgg agatccaatt tttaagtgta taatgtgtta aactactgat
     2221 tctaattgtt tgtgtatttt agattcacag tcccaaggct catttcaggc ccctcagtcc
     2281 tcacagtctg ttcatgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2341 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2401 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2521 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2581 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2641 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2701 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2761 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2821 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2881 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2941 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatcctg
     3121 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3481 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3541 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3601 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3661 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3721 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3781 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3841 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3901 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3961 gtcttgtcga tcaggatgat ctggacgaag aacatcaggg gctcgcgcca gccgaactgt
     4021 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4081 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4141 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4201 aacttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4261 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4321 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4441 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4501 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4561 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4621 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4681 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4741 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4801 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4861 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4921 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4981 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5041 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5101 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5161 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5221 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5281 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5341 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5401 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5461 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5521 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5581 tgattctgtg gataaccgta ttaccgcc
//
