LOCUS       sequence-444548-        6935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the short isoform of SYNJ1 (neuronal) with mCherry tagged 
            at the n-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6935)
  AUTHORS   Mohd Rafiq N, Fujise K, Rosenfeld MS, Xu P, De Camilli P
  TITLE     Parkinsonism Sac domain mutation in Synaptojanin-1 affects ciliary 
            properties in iPSC-derived dopaminergic neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Apr 23;121(17):e2318943121. doi: 
            10.1073/pnas.2318943121. Epub 2024 Apr 18.
  PUBMED    38635628
REFERENCE   2  (bases 1 to 6935)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-04-23"; volume: "121(17):e2318943121. 
            doi"; pages: " 10.1073/pnas.2318943121. Epub 2024 Apr 18"
COMMENT     Sequence Label: pmCherry-Synaptojanin-1-145
FEATURES             Location/Qualifiers
     source          1..6935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             619..636
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             646..666
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             667..1371
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(809..827)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1077..1096
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1399..1422
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3723..3844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3760..3779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3814..3833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3851..4306)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3988..4009)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4200..4219
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4333..4437
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4439..4796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4439..4630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4462..4482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4647..4782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4831..5625
                     /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             4831..5625
                     /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(4885..4904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5495..5514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5813..5832)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5857..5904
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6233..6821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6722..6741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatgggcca tcaccatcac catcacagcg gcagcgagaa cctgtacttc
      661 cagggcgtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
      721 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      781 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
      841 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
      901 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      961 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1021 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1081 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1141 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1201 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1261 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1321 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtccggactc
     1381 ggcggccacc gctggatcct ggaggtgctg ttccagggcc ccagatctat gaactacctg
     1441 cgccgccgcc tgagcgacag caacttcatg gccaacctgc ccaacggcta catgaccgac
     1501 ctgcagcgcc cccccccccc cccccccccc cccggcgccc acagccccgg cgccaccccc
     1561 ggccccggca ccgccaccgc cgagcgcagc agcggcgtgg cccccgccgc cagccccgcc
     1621 gcccccagcc ccggcagcag cggcggcggc ggcttcttca gcagcctgag caacgccgtg
     1681 aagcagacca ccgccgccgc cgccgccacc ttcagcgagc aggtgggcgg cggcagcggc
     1741 ggcgccggcc gcggcggcgc cgccagccgc gtgctgctgg tgatcgacga gccccacacc
     1801 gactgggcca agtacttcaa gggcaagaag atccacggcg agatcgacat caaggtggag
     1861 caggccgagt tcagcgacct gaacctggtg gcccacgcca acggcggctt cagcgtggac
     1921 atggaggtgc tgcgcaacgg cgtgaaggtg gtgcgcagcc tgaagcccga cttcgtgctg
     1981 atccgccagc acgccttcag catggcccgc aacggcgact accgcagcct ggtgatcggc
     2041 ctgcagtacg ccggcatccc cagcgtgaac agcctgcaca gcgtgtacaa cttctgcgac
     2101 aagccctggg tgttcgccca gatggtgcgc ctgcacaaga agctgggcac cgaggagttc
     2161 cccctgatcg accagacctt ctaccccaac cacaaggaga tgctgagcag caccacctac
     2221 cccgtggtgg tgaagatggg ccacgcccac agcggcatgg gcaaggtgaa ggtggacaac
     2281 cagcacgact tccaggacat cgccagcgtg gtggccctga ccaagaccta cgccaccgcc
     2341 gagcccttca tcgacgccaa gtacgacgtg cgcgtgcaga agatcggcca gaactacaag
     2401 gcctacatgc gcaccagcgt gagcggcaac tggaagacca acaccggcag cgccatgctg
     2461 gagcagatcg ccatgagcga ccgctacaag ctgtgggtgg acacctgcag cgagatattc
     2521 ggcggcctgg acatctgcgc cgtggaggcc ctgcacggca aggacggccg cgaccacatc
     2581 atcgaggtgg tgggcagcag catgcccctg atcggcgacc accaggacga ggacaagcag
     2641 ctgatcgtgg agctggtggt gaacaagatg gcccaggccc tgccccgcca gcgccagcgc
     2701 gacgccagcc ccggccgcgg cagccacggc cagaccccca gccccggcgc cctgcccctg
     2761 ggccgccaga ccagccagca gcccgccggc ccccccgccc agcagcgccc ccccccccag
     2821 ggcggccccc cccagcccgg ccccggcccc cagcgccagg gcccccccct gcagcagcgc
     2881 cccccccccc agggccagca gcacctgagc ggcctgggcc cccccgccgg cagccccctg
     2941 ccccagcgcc tgcccagccc caccagcgcc ccccagcagc ccgccagcca ggccgccccc
     3001 cccacccagg gccagggccg ccagagccgc cccgtggccg gcggccccgg cgcccccccc
     3061 gccgcccgcc cccccgccag ccccagcccc cagcgccagg ccggcccccc ccaggccacc
     3121 cgccagacca gcgtgagcgg ccccgccccc cccaaggcca gcggcgcccc ccccggcggc
     3181 cagcagcgcc agggcccccc ccagaagccc cccggccccg ccggccccac ccgccaggcc
     3241 agccaggccg gccccgtgcc ccgcaccggc ccccccacca cccagcagcc ccgccccagc
     3301 ggccccggcc ccgccggccg ccccaagccc cagctggccc agaagcccag ccaggacgtg
     3361 cccccccccg ccaccgccgc cgccggcggc cccccccacc cccagctgaa caagagccag
     3421 agcctgacca acgccttcaa cctgcccgag cccgcccccc cccgccccag cctgagccag
     3481 gacgaggtga aggccgagac catccgcagc ctgcgcaaga gcttcgccag cctgttcagc
     3541 gactgataag agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
     3601 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3661 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3721 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3781 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3841 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3901 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3961 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4021 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4081 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4141 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4201 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4261 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4321 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4381 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4441 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4621 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4681 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4801 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4861 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4921 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4981 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     5041 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5101 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5161 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5221 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5281 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5341 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5401 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5461 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5521 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5581 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5641 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5701 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5761 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5821 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5881 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5941 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6001 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6061 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     6121 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6181 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6241 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6301 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6361 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6421 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6481 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6541 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6601 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6661 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6721 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6781 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6841 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6901 ctgattctgt ggataaccgt attaccgcca tgcat
//
