LOCUS       sequence-489398-        5989 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Express RILPL1 in mammalian cells with mBaoJin tag (monomeric 
            StayGold).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5989)
  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 5989)
  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: mBaoJin-RILPL1
FEATURES             Location/Qualifiers
     source          1..5989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..81
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      149..158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             155..856
                     /codon_start=1
                     /product="bright and photostable monomeric green 
                     fluorescent protein derived from StayGold (Zhang et al., 
                     2024)"
                     /label=mBaoJin
                     /label=mStayGold
                     /note="/vntifkey=21"
                     /translation="MVSKGEEENMASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTTFGYGMKYYTKYPSGLKNWFREVMPGGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVPQIPRDDGVECPVTL
                     LYPLLSDKSKYVEAHQYTICKPLHNQPAPDVPYHWIRKQYKQSKDDAEERDHICQSETL
                     EAHLKGMDELYK"
     protein_bind    881..905
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2145..2169)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2319..2440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2356..2375)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2410..2429
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2447..2902)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2584..2605)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2796..2815
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2929..3033
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3035..3392
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3035..3226
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3058..3078)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3243..3378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3427..4221
                     /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             3427..4221
                     /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(3481..3500)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4091..4110
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4409..4428)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4453..4500
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4829..5417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5318..5337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5592..5895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5896..5989,1..110)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
       61 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      121 accgtcagat ccgctagcgc taccggtcgc caccatggtc tcaaagggag aggaggaaaa
      181 catggctagt acaccattta aatttcaact taaaggaacc atcaatggca aatcgtttac
      241 cgttgaaggc gaaggtgaag ggaactcaca tgaaggttct cataaaggaa aatatgtttg
      301 tacaagtgga aaactaccga tgtcatgggc agcacttggg acaacctttg gttatggaat
      361 gaaatattat accaaatatc ctagtggact gaagaactgg tttcgtgaag taatgcccgg
      421 aggctttacc tacgatcgtc atattcaata taaaggcgat gggagtatcc atgcaaaaca
      481 ccaacacttt atgaaaaatg ggacttatca caacattgta gaatttaccg gtcaggattt
      541 taaagaaaat agtccagtct taactggaga tatgaatgtc tcattaccga atgaagtccc
      601 acaaataccc agagatgatg gagtagaatg cccagtgacc ttgctttatc ctttattatc
      661 ggataaatca aaatacgtcg aggctcacca atatacaatc tgcaagcctc ttcataatca
      721 accagcacct gatgtcccat atcactggat tcgtaaacaa tacaaacaaa gcaaagatga
      781 tgccgaggaa cgcgatcata tctgtcaatc agagactctc gaagcacact taaagggcat
      841 ggacgagctg tataagtccg gactcagatc tcgagatcaa acaagtttgt acaaaaaagc
      901 aggctccgaa ttcgccctta tggaggagga gcgggggtcg gcgctggcgg ccgagtcggc
      961 gctggagaag aacgtggccg agctgaccgt catggacgtg tacgacatcg cgtcgcttgt
     1021 gggccacgag ttcgagcggg tcattgacca gcacggctgc gaggccatcg cgcgcctcat
     1081 gcccaaggtc gtgcgcgtcc tggagatcct ggaggtgctg gtcagccgcc accacgtcgc
     1141 gcccgagctg gacgagctgc gcctggagct ggaccgcctg cgcctggaga ggatggaccg
     1201 catcgagaag gagcgcaagc accagaagga gctggagctg gtggaggatg tgtggcgagg
     1261 ggaggcgcag gacctcctct cccagatcgc ccagctgcag gaggagaaca agcagctcat
     1321 gaccaacctc tcccacaagg atgtcaactt ctcagaggag gagttccaga agcatgaagg
     1381 catgtcagag cgggagcgac aggtgatgaa gaagctgaag gaggtggtgg acaaacaacg
     1441 cgacgagatc cgcgccaagg acagggagct gggcctgaaa aatgaggacg ttgaggcttt
     1501 acagcagcag cagacacggc tgatgaagat caaccatgac cttcggcacc gggtcacggt
     1561 ggtggaggcc caggggaaag ccctgatcga acagaaggtg gagctggagg cagacctgca
     1621 gaccaaggag caggagatgg gcagcctgcg agcagagctg gggaagttgc gagagaggct
     1681 gcagggggag cacagccaga atggggagga ggagcctgag acggagccgg tgggagagga
     1741 gagcatctcc gacgcagaga aggtggccat ggatctcaag gaccccaacc gcccccggtt
     1801 caccctgcag gagctgcggg acgtgctgca cgagaggaac gagctcaagt ccaaggtgtt
     1861 cttgctgcag gaggagctgg cttactataa gagtgaagaa atggaagagg aaaaccgaat
     1921 accccaaccc ccacccatcg cccacccgag gacgtccccc cagccggagt cgggcatcaa
     1981 gcgactgttt agcttcttct cccgagataa gaagcgcctg gccaacacac agagaaacgt
     2041 gcacatccag gagtcctttg gacagtgggc aaacacccac cgcgatgacg gttacacaga
     2101 gcaaggacag gaagccctgc agcatctgtg aaagggcgaa ttcgacccag ctttcttgta
     2161 caaagtggtt cgatggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     2221 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2281 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2341 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2401 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2461 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2521 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2581 gttccagttt ggaacaagag tccactatca aagaacgtgg actccaacgt caaagggcga
     2641 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2701 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2761 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2821 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2881 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2941 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3001 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3061 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3301 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3361 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3421 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3481 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3541 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3601 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3661 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3721 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3781 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3841 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3901 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3961 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4021 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4081 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4141 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4201 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4261 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4321 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4381 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4441 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4501 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4561 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4621 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4681 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4741 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4801 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4861 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4921 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4981 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5041 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5101 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5161 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5221 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5281 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5341 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5401 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5461 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5521 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     5581 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5641 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5701 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5761 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5821 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     5881 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     5941 gactcacggg gatttccaag tctccacccc attgacgtca atgggagtt
//
