LOCUS       sequence-444551-        6642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the full-length mouse INPP5E (Pharbin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6642)
  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 6642)
  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: EGFP-INPP5E
FEATURES             Location/Qualifiers
     source          1..6642
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3430..3551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3467..3486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3521..3540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3558..4013)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3695..3716)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3907..3926
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4040..4144
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4146..4503
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4146..4337
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4169..4189)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4354..4489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4538..5332
                     /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             4538..5332
                     /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(4592..4611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5520..5539)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5564..5611
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5940..6528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6429..6448
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggaatgcc atccaagtca gcttgcctgc gtcacactga ggcacctggg
     1381 cagctagaag gcagaatgct ccagggacag cctccgaaca cggaaaagaa gctcatccca
     1441 acaccaggct tcttaccagc ctctgactct cagggttcag agacaaaccc catgcctcct
     1501 ttcagcattc cagcaaaaac aagcaaccag aacccacaaa ctaaggcaaa tctcatcacc
     1561 ccacagccac ccattagacc caagttggag cgaaccctgt ccctcgatga caagggctgg
     1621 agaaggaggc gtttccgggg tagtcaggag gatctgactg tccagaatgg ggccagtccc
     1681 tgcaggggct ccttgcaaga ctcagtagcc cagtcccctg cctacagccg tcccctgccc
     1741 tgcctcagca catccttgca ggagatacct aagtcccgaa gggccacagg cagtgaggga
     1801 gggagcccat ccttgtggag tgactgtctt tctggaatga tcagcacctc cttggacctc
     1861 ctgcacagag atgctgcctc aggtgggccc ccctccaggt tggcaagttt gcatgcctca
     1921 cacacaccac cagctatgga cctcagcata gcctccagct ccctgagaac agcaaacaag
     1981 gttgaccccg agcacacgga ctacaagctc cgcatgcaga ccagactggt cagggcccac
     2041 agcaacctgg gccctagcag accccggagc cccttggctg gtgacgatca ctccattcac
     2101 tcagccagat ctttcagcct tctggccccc atccgcacca aggacatcag aagcaggagc
     2161 tatctggagg gaagtcttct ggccagtggg gccctgctag gagcagaaga gctggccagg
     2221 tacttcccag accgaaacat ggctctcttc gtggctacct ggaacatgca gggccagaag
     2281 gagctcccag cgagcctgga tgagtttctg ctccccaccg aggctgacta cactcaggac
     2341 ctgtatgtca ttggaattca ggagggctgc tctgacaggc gggagtggga gacacgcctg
     2401 caggagacac tgggccctca gtatgtactg ctgtcatcag cagcacatgg ggtcctgtac
     2461 atgtccctgt ttatccgtag ggacctcatc tggttctgct cagaggtcga gtactccaca
     2521 gtaactacac gcatcgtgtc tcagatcaag accaaggggg ccctgggcgt cagcttcacc
     2581 ttttttggca cctccttcct cttcatcaca tctcacttca cctctggaga tgggaaggta
     2641 gcagagcggc tactggacta cagcagaacc atccaagccc tagccctgcc ccggaatgtg
     2701 ccagacacaa atccctaccg ctctagtgca ggggatgtca ctacccggtt tgatgaggtc
     2761 ttctggtttg gggacttcaa cttccgcctg agtggtggac gagtggctgt ggaggccttc
     2821 ctgaagcaga aacctgaggt ggatgtgctg gctctcctcc aacacgacca gctcacccgg
     2881 gagatgaaga aagggtccat cttcaggggc tttgaggagg cagagattca ctttcttcca
     2941 tcctacaagt ttgacattgg gaaggacacc tacgacagca cctccaagca aaggacaccc
     3001 tcctacacag accgagtcct atacaaaagc cgtcacaagg gtgacatctg tcccatgaag
     3061 tattcctctt gtcctgggat caagacttca gaccaccgtc ctgtgtatgg cttgttccag
     3121 gtgaaagtga ggccaggacg agacaacatc ccattagctg ccggcaagtt tgaccgagaa
     3181 ttgtacttga taggaattaa aaggcggatt tcaaaggaga ttcagagaca ggaagcactg
     3241 aagagccaga gctccagtgc agtttgcacc gtgtcctgag tcgacggtac cgcgggcccg
     3301 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     3361 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3421 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3541 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3601 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3661 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3721 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3781 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3841 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3901 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3961 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4021 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4081 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4141 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4261 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4321 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4381 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4441 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4501 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5401 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5461 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5521 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5581 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5641 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5701 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5761 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5821 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5881 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5941 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6001 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6061 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6121 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6181 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6241 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6301 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6361 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6421 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6481 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6541 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6601 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
