LOCUS       sequence-489397-        8743 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Express JIP4 in mammalian cells with mBaoJin tag (monomeric 
            StayGold).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8743)
  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 8743)
  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-JIP4
FEATURES             Location/Qualifiers
     source          1..8743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1321
                     /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    1346..1370
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5364..5388)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5538..5659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5575..5594)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5629..5648
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5666..6121)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5803..5824)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6015..6034
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6148..6252
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6254..6611
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6254..6445
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6277..6297)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6462..6597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6646..7440
                     /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             6646..7440
                     /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(6700..6719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7310..7329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7628..7647)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7672..7719
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8048..8636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8537..8556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtctcaaa gggagaggag gaaaacatgg ctagtacacc
      661 atttaaattt caacttaaag gaaccatcaa tggcaaatcg tttaccgttg aaggcgaagg
      721 tgaagggaac tcacatgaag gttctcataa aggaaaatat gtttgtacaa gtggaaaact
      781 accgatgtca tgggcagcac ttgggacaac ctttggttat ggaatgaaat attataccaa
      841 atatcctagt ggactgaaga actggtttcg tgaagtaatg cccggaggct ttacctacga
      901 tcgtcatatt caatataaag gcgatgggag tatccatgca aaacaccaac actttatgaa
      961 aaatgggact tatcacaaca ttgtagaatt taccggtcag gattttaaag aaaatagtcc
     1021 agtcttaact ggagatatga atgtctcatt accgaatgaa gtcccacaaa tacccagaga
     1081 tgatggagta gaatgcccag tgaccttgct ttatccttta ttatcggata aatcaaaata
     1141 cgtcgaggct caccaatata caatctgcaa gcctcttcat aatcaaccag cacctgatgt
     1201 cccatatcac tggattcgta aacaatacaa acaaagcaaa gatgatgccg aggaacgcga
     1261 tcatatctgt caatcagaga ctctcgaagc acacttaaag ggcatggacg agctgtataa
     1321 gtccggactc agatctcgag atcaaacaag tttgtacaaa aaagcaggct ccgaattcgc
     1381 ccttatggag ctggaggacg gtgtggtgta tcaggaggag cccggcggct ccggggccgt
     1441 gatgtcggag cgggtgtccg gcctggccgg ctccatctac cgcgagttcg agcggcttat
     1501 cgggcgctat gacgaggagg tggtcaaaga gctgatgccg ctggtggtgg ctgtgctgga
     1561 gaacctggac tcggtgttcg cgcaggacca ggagcaccag gtggagctgg agctgctgcg
     1621 ggacgacaac gagcagctca tcacccagta cgagcgggag aaggcgctgc gcaagcacgc
     1681 tgaggagaaa ttcattgaat ttgaagactc tcaagaacag gaaaaaaagg acttacagac
     1741 ccgagtggaa tctttagaat ctcaaacaag acaacttgag ctgaaagcga aaaactatgc
     1801 tgaccagatt agcagacttg aagaaagaga agcagaactg aagaaggaat ataatgcatt
     1861 acatcaaaga cacactgaga tgatccataa ttatatggaa catttagaaa gaacaaaact
     1921 tcatcagctc tcagggagtg atcaactaga atccacagct catagtagaa ttagaaaaga
     1981 acgccctata tcattaggaa ttttcccatt acctgctgga gatggattgc ttacacctga
     2041 tgctcagaaa ggaggagaga cccctggatc tgagcaatgg aaatttcagg aattaagtca
     2101 accacgttct cataccagcc tgaaggtcag caatagtcct gaacctcaga aggctgtaga
     2161 acaggaggat gagctttctg atgttagcca aggcggatct aaagctacca ctccagcatc
     2221 aacagctaat tcagatgtgg caacaattcc tactgatact cccttaaagg aagaaaacga
     2281 aggatttgtg aaggttacag atgcgccaaa taaatcagag ataagcaaac acattgaagt
     2341 acaggtagcc caggaaacta gaaatgtatc tactggctct gctgaaaatg aagaaaagtc
     2401 agaagttcaa gcaatcatcg aatctactcc tgagctggat atggacaaag atctcagtgg
     2461 atataaaggt tcaagcactc ccaccaaagg catagagaac aaagcttttg atcgcaatac
     2521 agaatctctc tttgaagaac tgtcttcagc tggctcaggc ctaataggag atgtggatga
     2581 aggagcagat ttactaggaa tgggtcggga agttgagaat cttatattag aaaatacaca
     2641 actgttggaa accaaaaatg ctttgaacat agtgaagaat gatttgatag caaaagtgga
     2701 tgaactgacc tgtgagaaag atgtgctgca aggggaattg gaggctgtga agcaagccaa
     2761 actgaaacta gaggaaaaga acagagaatt ggaggaagag cttaggaaag ctcgggcaga
     2821 agctgaagat gcaaggcaaa aagcaaaaga tgacgatgat agtgatattc ccacagccca
     2881 gaggaaacgg tttactagag tagaaatggc ccgtgttctc atggagcgaa accagtataa
     2941 agagagattg atggagcttc aggaagctgt tcgatggaca gagatgattc gggcatcacg
     3001 agaaaatcca gccatgcagg aaaaaaaaag gtcaagcatt tggcagtttt tcagccgact
     3061 tttcagctcc tcaagtaaca cgactaagaa gcctgaacca cctgttaatc tgaagtacaa
     3121 tgcacccacg tctcatgtta ctccgtccgt caagaaaaga agcagcacct tatctcagct
     3181 ccctggggat aagtccaaag cctttgattt ccttagtgaa gaaactgaag ctagtttagc
     3241 ctcacgcaga gaacaaaaga gagagcagta tcgtcaggta aaagcacatg ttcagaagga
     3301 agacggtaga gtgcaggctt ttggctggag tctgcctcag aagtacaaac aggtaaccaa
     3361 tggtcaaggt gaaaataaga tgaaaaattt acctgtgcct gtctatctca gacctctgga
     3421 tgaaaaagat acatcaatga agctgtggtg tgctgttgga gtcaatttat ctggtgggaa
     3481 gaccagagat ggtggttctg ttgttggagc aagtgtattt tacaaggatg ttgctggttt
     3541 ggatacagaa ggcagtaaac agcgaagtgc ctctcagagt agtttagata agttagatca
     3601 ggaacttaag gaacagcaga aggagttaaa aaatcaagaa gaattatcca gtctagtttg
     3661 gatctgtacc agcactcatt cggctacaaa agttcttatt attgatgctg ttcaacctgg
     3721 caacatccta gacagtttca ctgtttgcaa ctctcatgtt ctgtgcattg caagtgtgcc
     3781 aggtgcacga gaaacagact accctgcagg agaagatctt tcagaatctg gtcaggtaga
     3841 caaagcatct ttatgtggaa gtatgacaag caacagctca gcagagacag acagcctgtt
     3901 aggaggcatc acagtggttg gttgttctgc agaaggtgtg acgggagctg ccacttcccc
     3961 tagtacaaat ggtgcttctc cagtgatgga taaaccacca gaaatggaag cagaaaatag
     4021 tgaggttgat gaaaatgttc caacagcaga agaagcaact gaagctacag aagggaatgc
     4081 ggggtcagct gaagacacag tggacatctc ccaaactggc gtctacacag agcatgtctt
     4141 tacagatcct ttgggagttc agatcccaga agacctctcc ccagtgtatc agtcgagcaa
     4201 tgactcagat gcatataaag atcaaatatc agtactgcca aatgaacaag acttggtgag
     4261 agaagaagcc cagaaaatga gtagtctttt accaactatg tggcttggag ctcaaaatgg
     4321 ctgtttgtat gtccattcat ctgtagccca gtggaggaaa tgtctccatt ccattaaact
     4381 taaagattcg attctcagta ttgtacacgt gaagggaatc gtgttagtag ccctggctga
     4441 cggcaccctt gcaatctttc acagaggagt ggatgggcag tgggatttgt caaactatca
     4501 cctcttagac cttggacggc ctcatcattc catccgttgc atgactgtgg tacatgacaa
     4561 agtctggtgt ggctatagga acaaaatcta tgtggtgcag ccaaaggcca tgaaaataga
     4621 gaaatctttt gatgcacatc ccaggaagga gagccaagtg cgacagcttg cgtgggtggg
     4681 ggatggcgtg tgggtctcca ttcgcttgga ttctacgctc cgtctctatc atgcacacac
     4741 ttatcaacat ctacaggatg tggacattga gccttatgta agcaaaatgt taggtactgg
     4801 aaaactgggc ttctcttttg tgagaattac agctcttatg gtgtcttgta atcgtttgtg
     4861 ggtggggaca ggaaatggtg tcattatctc catcccattg acagaaacaa ataaaacctc
     4921 aggtgtacca ggaaatcgtc ctggaagtgt aatccgtgta tatggtgatg aaaacagtga
     4981 taaagtgact ccagggacat ttatacccta ttgttcaatg gcacatgcac agctttgctt
     5041 ccatgggcac cgggatgctg tgaaattctt tgtggcagtc ccaggtcaag tcatcagccc
     5101 acaaagtagc agtagtggca cggatctgac gggtgacaaa gcagggccat ctgcacagga
     5161 gcctggtagt cagacgccct tgaagtctat gcttgtcatc agtggaggag agggctacat
     5221 cgacttccga atgggtgatg aaggtggaga atcagaactt cttggagagg atcttccact
     5281 tgaaccttct gtcaccaaag cagaaaggag tcacttgata gtgtggcaag tgatgtatgg
     5341 caatgagtga aagggcgaat tcgacccagc tttcttgtac aaagtggttc gatggtaccg
     5401 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     5461 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5521 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5581 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5641 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     5701 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5761 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5821 ccactatcaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5881 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5941 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     6001 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6061 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6121 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6181 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6241 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     6301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6361 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6421 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6481 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6541 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6601 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6661 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6721 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6781 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6841 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6901 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6961 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     7021 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     7081 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     7141 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     7201 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     7261 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     7321 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7381 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7441 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     7501 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     7561 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     7621 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     7681 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     7741 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     7801 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7861 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     7921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8161 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8701 ttcctgcgtt atcccctgat tctgtggata accgtattac cgc
//
