LOCUS       sequence-324159-        8749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses JIP4 in mammalian cells with a mNeonGreen tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8749)
  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 8749)
  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: mNeonGreen-JIP4
FEATURES             Location/Qualifiers
     source          1..8749
                     /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..1327
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      875..884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1352..1376
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5370..5394)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5544..5665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5581..5600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5635..5654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5672..6127)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5809..5830)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6021..6040
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6154..6258
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6260..6617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6260..6451
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6283..6303)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6468..6603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6652..7446
                     /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             6652..7446
                     /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(6706..6725)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7316..7335
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7634..7653)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7678..7725
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8054..8642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8543..8562
                     /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 tggtgagcaa gggcgaggag gataacatgg cctctctccc
      661 agcgacacat gagttacaca tctttggctc catcaacggt gtggactttg acatggtggg
      721 tcagggcacc ggcaatccaa atgatggtta tgaggagtta aacctgaagt ccaccaaggg
      781 tgacctccag ttctccccct ggattctggt ccctcatatc gggtatggct tccatcagta
      841 cctgccctac cctgacggga tgtcgccttt ccaggccgcc atggtagatg gctccggcta
      901 ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc cttactgtta actaccgcta
      961 cacctacgag ggaagccaca tcaaaggaga ggcccaggtg aaggggactg gtttccctgc
     1021 tgacggtcct gtgatgacca actcgctgac cgctgcggac tggtgcaggt cgaagaagac
     1081 ttaccccaac gacaaaacca tcatcagtac ctttaagtgg agttacacca ctggaaatgg
     1141 caagcgctac cggagcactg cgcggaccac ctacaccttt gccaagccaa tggcggctaa
     1201 ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc actccaagac
     1261 cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca tggacgagct
     1321 gtacaagtcc ggactcagat ctcgagatca aacaagtttg tacaaaaaag caggctccga
     1381 attcgccctt atggagctgg aggacggtgt ggtgtatcag gaggagcccg gcggctccgg
     1441 ggccgtgatg tcggagcggg tgtccggcct ggccggctcc atctaccgcg agttcgagcg
     1501 gcttatcggg cgctatgacg aggaggtggt caaagagctg atgccgctgg tggtggctgt
     1561 gctggagaac ctggactcgg tgttcgcgca ggaccaggag caccaggtgg agctggagct
     1621 gctgcgggac gacaacgagc agctcatcac ccagtacgag cgggagaagg cgctgcgcaa
     1681 gcacgctgag gagaaattca ttgaatttga agactctcaa gaacaggaaa aaaaggactt
     1741 acagacccga gtggaatctt tagaatctca aacaagacaa cttgagctga aagcgaaaaa
     1801 ctatgctgac cagattagca gacttgaaga aagagaagca gaactgaaga aggaatataa
     1861 tgcattacat caaagacaca ctgagatgat ccataattat atggaacatt tagaaagaac
     1921 aaaacttcat cagctctcag ggagtgatca actagaatcc acagctcata gtagaattag
     1981 aaaagaacgc cctatatcat taggaatttt cccattacct gctggagatg gattgcttac
     2041 acctgatgct cagaaaggag gagagacccc tggatctgag caatggaaat ttcaggaatt
     2101 aagtcaacca cgttctcata ccagcctgaa ggtcagcaat agtcctgaac ctcagaaggc
     2161 tgtagaacag gaggatgagc tttctgatgt tagccaaggc ggatctaaag ctaccactcc
     2221 agcatcaaca gctaattcag atgtggcaac aattcctact gatactccct taaaggaaga
     2281 aaacgaagga tttgtgaagg ttacagatgc gccaaataaa tcagagataa gcaaacacat
     2341 tgaagtacag gtagcccagg aaactagaaa tgtatctact ggctctgctg aaaatgaaga
     2401 aaagtcagaa gttcaagcaa tcatcgaatc tactcctgag ctggatatgg acaaagatct
     2461 cagtggatat aaaggttcaa gcactcccac caaaggcata gagaacaaag cttttgatcg
     2521 caatacagaa tctctctttg aagaactgtc ttcagctggc tcaggcctaa taggagatgt
     2581 ggatgaagga gcagatttac taggaatggg tcgggaagtt gagaatctta tattagaaaa
     2641 tacacaactg ttggaaacca aaaatgcttt gaacatagtg aagaatgatt tgatagcaaa
     2701 agtggatgaa ctgacctgtg agaaagatgt gctgcaaggg gaattggagg ctgtgaagca
     2761 agccaaactg aaactagagg aaaagaacag agaattggag gaagagctta ggaaagctcg
     2821 ggcagaagct gaagatgcaa ggcaaaaagc aaaagatgac gatgatagtg atattcccac
     2881 agcccagagg aaacggttta ctagagtaga aatggcccgt gttctcatgg agcgaaacca
     2941 gtataaagag agattgatgg agcttcagga agctgttcga tggacagaga tgattcgggc
     3001 atcacgagaa aatccagcca tgcaggaaaa aaaaaggtca agcatttggc agtttttcag
     3061 ccgacttttc agctcctcaa gtaacacgac taagaagcct gaaccacctg ttaatctgaa
     3121 gtacaatgca cccacgtctc atgttactcc gtccgtcaag aaaagaagca gcaccttatc
     3181 tcagctccct ggggataagt ccaaagcctt tgatttcctt agtgaagaaa ctgaagctag
     3241 tttagcctca cgcagagaac aaaagagaga gcagtatcgt caggtaaaag cacatgttca
     3301 gaaggaagac ggtagagtgc aggcttttgg ctggagtctg cctcagaagt acaaacaggt
     3361 aaccaatggt caaggtgaaa ataagatgaa aaatttacct gtgcctgtct atctcagacc
     3421 tctggatgaa aaagatacat caatgaagct gtggtgtgct gttggagtca atttatctgg
     3481 tgggaagacc agagatggtg gttctgttgt tggagcaagt gtattttaca aggatgttgc
     3541 tggtttggat acagaaggca gtaaacagcg aagtgcctct cagagtagtt tagataagtt
     3601 agatcaggaa cttaaggaac agcagaagga gttaaaaaat caagaagaat tatccagtct
     3661 agtttggatc tgtaccagca ctcattcggc tacaaaagtt cttattattg atgctgttca
     3721 acctggcaac atcctagaca gtttcactgt ttgcaactct catgttctgt gcattgcaag
     3781 tgtgccaggt gcacgagaaa cagactaccc tgcaggagaa gatctttcag aatctggtca
     3841 ggtagacaaa gcatctttat gtggaagtat gacaagcaac agctcagcag agacagacag
     3901 cctgttagga ggcatcacag tggttggttg ttctgcagaa ggtgtgacgg gagctgccac
     3961 ttcccctagt acaaatggtg cttctccagt gatggataaa ccaccagaaa tggaagcaga
     4021 aaatagtgag gttgatgaaa atgttccaac agcagaagaa gcaactgaag ctacagaagg
     4081 gaatgcgggg tcagctgaag acacagtgga catctcccaa actggcgtct acacagagca
     4141 tgtctttaca gatcctttgg gagttcagat cccagaagac ctctccccag tgtatcagtc
     4201 gagcaatgac tcagatgcat ataaagatca aatatcagta ctgccaaatg aacaagactt
     4261 ggtgagagaa gaagcccaga aaatgagtag tcttttacca actatgtggc ttggagctca
     4321 aaatggctgt ttgtatgtcc attcatctgt agcccagtgg aggaaatgtc tccattccat
     4381 taaacttaaa gattcgattc tcagtattgt acacgtgaag ggaatcgtgt tagtagccct
     4441 ggctgacggc acccttgcaa tctttcacag aggagtggat gggcagtggg atttgtcaaa
     4501 ctatcacctc ttagaccttg gacggcctca tcattccatc cgttgcatga ctgtggtaca
     4561 tgacaaagtc tggtgtggct ataggaacaa aatctatgtg gtgcagccaa aggccatgaa
     4621 aatagagaaa tcttttgatg cacatcccag gaaggagagc caagtgcgac agcttgcgtg
     4681 ggtgggggat ggcgtgtggg tctccattcg cttggattct acgctccgtc tctatcatgc
     4741 acacacttat caacatctac aggatgtgga cattgagcct tatgtaagca aaatgttagg
     4801 tactggaaaa ctgggcttct cttttgtgag aattacagct cttatggtgt cttgtaatcg
     4861 tttgtgggtg gggacaggaa atggtgtcat tatctccatc ccattgacag aaacaaataa
     4921 aacctcaggt gtaccaggaa atcgtcctgg aagtgtaatc cgtgtatatg gtgatgaaaa
     4981 cagtgataaa gtgactccag ggacatttat accctattgt tcaatggcac atgcacagct
     5041 ttgcttccat gggcaccggg atgctgtgaa attctttgtg gcagtcccag gtcaagtcat
     5101 cagcccacaa agtagcagta gtggcacgga tctgacgggt gacaaagcag ggccatctgc
     5161 acaggagcct ggtagtcaga cgcccttgaa gtctatgctt gtcatcagtg gaggagaggg
     5221 ctacatcgac ttccgaatgg gtgatgaagg tggagaatca gaacttcttg gagaggatct
     5281 tccacttgaa ccttctgtca ccaaagcaga aaggagtcac ttgatagtgt ggcaagtgat
     5341 gtatggcaat gagtgaaagg gcgaattcga cccagctttc ttgtacaaag tggttcgatg
     5401 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     5461 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5521 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5581 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5641 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     5701 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5761 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5821 aagagtccac tatcaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5881 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5941 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6001 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6061 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6121 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6181 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6241 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6361 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6481 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6541 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6601 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     6661 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     6721 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6781 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     6841 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6901 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6961 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     7021 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     7081 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     7141 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     7201 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7261 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7321 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7381 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7441 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     7501 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7561 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     7621 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     7681 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     7741 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     7801 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     7861 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     7921 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7981 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8041 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8101 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8161 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8221 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8281 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8341 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8401 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8461 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8521 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8581 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8641 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8701 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgc
//
