LOCUS       sequence-324323-        8362 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses JIP4 in mammalian cells with a 2xmyc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8362)
  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 8362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-JIP4
FEATURES             Location/Qualifiers
     source          1..8362
                     /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(4790..4814)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4863..4882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4864..4882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5156..5277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5193..5212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5247..5266
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5284..5739)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5421..5442)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5633..5652
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5766..5870
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5872..6229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5872..6063
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5895..5915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6080..6215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6264..7058
                     /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             6264..7058
                     /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(6318..6337)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6928..6947
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7246..7265)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7290..7337
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7666..8254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8155..8174
                     /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 atggagctgg aggacggtgt ggtgtatcag
      841 gaggagcccg gcggctccgg ggccgtgatg tcggagcggg tgtccggcct ggccggctcc
      901 atctaccgcg agttcgagcg gcttatcggg cgctatgacg aggaggtggt caaagagctg
      961 atgccgctgg tggtggctgt gctggagaac ctggactcgg tgttcgcgca ggaccaggag
     1021 caccaggtgg agctggagct gctgcgggac gacaacgagc agctcatcac ccagtacgag
     1081 cgggagaagg cgctgcgcaa gcacgctgag gagaaattca ttgaatttga agactctcaa
     1141 gaacaggaaa aaaaggactt acagacccga gtggaatctt tagaatctca aacaagacaa
     1201 cttgagctga aagcgaaaaa ctatgctgac cagattagca gacttgaaga aagagaagca
     1261 gaactgaaga aggaatataa tgcattacat caaagacaca ctgagatgat ccataattat
     1321 atggaacatt tagaaagaac aaaacttcat cagctctcag ggagtgatca actagaatcc
     1381 acagctcata gtagaattag aaaagaacgc cctatatcat taggaatttt cccattacct
     1441 gctggagatg gattgcttac acctgatgct cagaaaggag gagagacccc tggatctgag
     1501 caatggaaat ttcaggaatt aagtcaacca cgttctcata ccagcctgaa ggtcagcaat
     1561 agtcctgaac ctcagaaggc tgtagaacag gaggatgagc tttctgatgt tagccaaggc
     1621 ggatctaaag ctaccactcc agcatcaaca gctaattcag atgtggcaac aattcctact
     1681 gatactccct taaaggaaga aaacgaagga tttgtgaagg ttacagatgc gccaaataaa
     1741 tcagagataa gcaaacacat tgaagtacag gtagcccagg aaactagaaa tgtatctact
     1801 ggctctgctg aaaatgaaga aaagtcagaa gttcaagcaa tcatcgaatc tactcctgag
     1861 ctggatatgg acaaagatct cagtggatat aaaggttcaa gcactcccac caaaggcata
     1921 gagaacaaag cttttgatcg caatacagaa tctctctttg aagaactgtc ttcagctggc
     1981 tcaggcctaa taggagatgt ggatgaagga gcagatttac taggaatggg tcgggaagtt
     2041 gagaatctta tattagaaaa tacacaactg ttggaaacca aaaatgcttt gaacatagtg
     2101 aagaatgatt tgatagcaaa agtggatgaa ctgacctgtg agaaagatgt gctgcaaggg
     2161 gaattggagg ctgtgaagca agccaaactg aaactagagg aaaagaacag agaattggag
     2221 gaagagctta ggaaagctcg ggcagaagct gaagatgcaa ggcaaaaagc aaaagatgac
     2281 gatgatagtg atattcccac agcccagagg aaacggttta ctagagtaga aatggcccgt
     2341 gttctcatgg agcgaaacca gtataaagag agattgatgg agcttcagga agctgttcga
     2401 tggacagaga tgattcgggc atcacgagaa aatccagcca tgcaggaaaa aaaaaggtca
     2461 agcatttggc agtttttcag ccgacttttc agctcctcaa gtaacacgac taagaagcct
     2521 gaaccacctg ttaatctgaa gtacaatgca cccacgtctc atgttactcc gtccgtcaag
     2581 aaaagaagca gcaccttatc tcagctccct ggggataagt ccaaagcctt tgatttcctt
     2641 agtgaagaaa ctgaagctag tttagcctca cgcagagaac aaaagagaga gcagtatcgt
     2701 caggtaaaag cacatgttca gaaggaagac ggtagagtgc aggcttttgg ctggagtctg
     2761 cctcagaagt acaaacaggt aaccaatggt caaggtgaaa ataagatgaa aaatttacct
     2821 gtgcctgtct atctcagacc tctggatgaa aaagatacat caatgaagct gtggtgtgct
     2881 gttggagtca atttatctgg tgggaagacc agagatggtg gttctgttgt tggagcaagt
     2941 gtattttaca aggatgttgc tggtttggat acagaaggca gtaaacagcg aagtgcctct
     3001 cagagtagtt tagataagtt agatcaggaa cttaaggaac agcagaagga gttaaaaaat
     3061 caagaagaat tatccagtct agtttggatc tgtaccagca ctcattcggc tacaaaagtt
     3121 cttattattg atgctgttca acctggcaac atcctagaca gtttcactgt ttgcaactct
     3181 catgttctgt gcattgcaag tgtgccaggt gcacgagaaa cagactaccc tgcaggagaa
     3241 gatctttcag aatctggtca ggtagacaaa gcatctttat gtggaagtat gacaagcaac
     3301 agctcagcag agacagacag cctgttagga ggcatcacag tggttggttg ttctgcagaa
     3361 ggtgtgacgg gagctgccac ttcccctagt acaaatggtg cttctccagt gatggataaa
     3421 ccaccagaaa tggaagcaga aaatagtgag gttgatgaaa atgttccaac agcagaagaa
     3481 gcaactgaag ctacagaagg gaatgcgggg tcagctgaag acacagtgga catctcccaa
     3541 actggcgtct acacagagca tgtctttaca gatcctttgg gagttcagat cccagaagac
     3601 ctctccccag tgtatcagtc gagcaatgac tcagatgcat ataaagatca aatatcagta
     3661 ctgccaaatg aacaagactt ggtgagagaa gaagcccaga aaatgagtag tcttttacca
     3721 actatgtggc ttggagctca aaatggctgt ttgtatgtcc attcatctgt agcccagtgg
     3781 aggaaatgtc tccattccat taaacttaaa gattcgattc tcagtattgt acacgtgaag
     3841 ggaatcgtgt tagtagccct ggctgacggc acccttgcaa tctttcacag aggagtggat
     3901 gggcagtggg atttgtcaaa ctatcacctc ttagaccttg gacggcctca tcattccatc
     3961 cgttgcatga ctgtggtaca tgacaaagtc tggtgtggct ataggaacaa aatctatgtg
     4021 gtgcagccaa aggccatgaa aatagagaaa tcttttgatg cacatcccag gaaggagagc
     4081 caagtgcgac agcttgcgtg ggtgggggat ggcgtgtggg tctccattcg cttggattct
     4141 acgctccgtc tctatcatgc acacacttat caacatctac aggatgtgga cattgagcct
     4201 tatgtaagca aaatgttagg tactggaaaa ctgggcttct cttttgtgag aattacagct
     4261 cttatggtgt cttgtaatcg tttgtgggtg gggacaggaa atggtgtcat tatctccatc
     4321 ccattgacag aaacaaataa aacctcaggt gtaccaggaa atcgtcctgg aagtgtaatc
     4381 cgtgtatatg gtgatgaaaa cagtgataaa gtgactccag ggacatttat accctattgt
     4441 tcaatggcac atgcacagct ttgcttccat gggcaccggg atgctgtgaa attctttgtg
     4501 gcagtcccag gtcaagtcat cagcccacaa agtagcagta gtggcacgga tctgacgggt
     4561 gacaaagcag ggccatctgc acaggagcct ggtagtcaga cgcccttgaa gtctatgctt
     4621 gtcatcagtg gaggagaggg ctacatcgac ttccgaatgg gtgatgaagg tggagaatca
     4681 gaacttcttg gagaggatct tccacttgaa ccttctgtca ccaaagcaga aaggagtcac
     4741 ttgatagtgt ggcaagtgat gtatggcaat gagtgaaagg gcgaattcga cccagctttc
     4801 ttgtacaaag tggtgatcgg taccttaatt aattaaggta ccaggtaagt gtacccaatt
     4861 cgccctatag tgagtcgtat tacaattcac tcgatcgccc ttcccaacag ttgcgcagcc
     4921 tgaatggcga atggagatcc aatttttaag tgtataatgt gttaaactac tgattctaat
     4981 tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca gtcctcacag
     5041 tctgttcatg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5101 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     5281 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5341 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5401 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5461 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5521 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5581 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5641 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5701 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     5761 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5821 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat cctgaggcgg
     5881 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6061 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     6241 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     6301 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6361 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     6421 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     6481 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     6541 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     6601 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6661 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6721 tcgatcagga tgatctggac gaagaacatc aggggctcgc gccagccgaa ctgttcgcca
     6781 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6841 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6901 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagaacttg
     6961 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     7021 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     7081 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     7141 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     7201 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     7261 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     7321 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     7381 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     7441 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     7501 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7801 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8341 tgtggataac cgtattaccg cc
//
