LOCUS       sequence-182295-        7961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cilia-membrane-targeted phosphatidylinositol 4-phosphate 5-kinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7961)
  AUTHORS   Phua SC, Chiba S, Suzuki M, Su E, Roberson EC, Pusapati GV, Setou M,
            Rohatgi R, Reiter JF, Ikegami K, Inoue T
  TITLE     Dynamic Remodeling of Membrane Composition Drives Cell Cycle through
            Primary Cilia Excision.
  JOURNAL   Cell. 2017 Jan 12;168(1-2):264-279.e15. doi: 
            10.1016/j.cell.2016.12.032. Epub 2017 Jan 12.
  PUBMED    28086093
REFERENCE   2  (bases 1 to 7961)
  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: "Cell."; 
            date: "2017-01-12"; volume: "168(1-2):264-279.e15. doi"; pages: " 
            10.1016/j.cell.2016.12.032. Epub 2017 Jan 12"
COMMENT     Sequence Label: 5HT6-YFP-PIPK
FEATURES             Location/Qualifiers
     source          1..7961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        90..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1982..1991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1988..1990,1991..1993,1994..2704)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2033..2054)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2294..2313)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2641..2662
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4778..4899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4815..4834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4869..4888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4906..5361)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5043..5064)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5255..5274
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5388..5492
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5494..5851
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5494..5685
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5517..5537)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5702..5837
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5886..6680
                     /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             5886..6680
                     /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(5940..5959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6550..6569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6868..6887)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6912..6959
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7288..7876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7777..7796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      601 ttagtgaacc gtcagatccg ctagcgccac catggttcca gagcccggcc ctgtcaacag
      661 tagcacccca gcctggggcc ccgggccacc gccagcccca gggggcagcg gctgggtggc
      721 cgctgcgctg tgcgtggtca tcgtgctgac ggcagccgcc aattcgctgc tgatcgcgct
      781 catctgcacg cagcccgcgc tgcgcaacac gtctaacttc ttcctggtgt cgctcttcac
      841 gtcggacctg atggtgggat tggtggtgat gcccccagcc atgctgaacg cgctgtatgg
      901 gcgctgggta ctagctcgcg gcctctgtct gctttggacc gccttcgacg tgatgtgctg
      961 cagcgcctcc attctcaacc tctgcctcat cagcctggac cgctacctgc tcatcctctc
     1021 gccgctgcgc tacaagctgc gcatgacagc cccgcgtgcc ctggcgctca tcctgggtgc
     1081 ctggagcctc gccgcgctgg cctccttcct gcccctcttg ctgggctggc atgaactggg
     1141 caaagctcga acatctgccc cgggccagtg ccgcctattg gccagcctgc cttatgtcct
     1201 cgtggcgtcc ggcgtcacct ttttcctgcc ttcgggtgcc atctgcttca cctactgcag
     1261 gatcttgctg gctgcccgca agcaggcggt gcaagtggcc tcgctcacca cgggcacggc
     1321 tacggctggc caggccttgg aaaccttgca ggtgcccagg acaccacgcc cagggatgga
     1381 gtctgctgac agtaggcgtc tgaccaccaa gcatagcagg aaggccttga aggccagcct
     1441 gaccctcggc atcctgctga gcatgttctt tgtcacctgg ctgcccttct ttgtggccag
     1501 catagctcag gccgtatgtg actgcatctc tccaggtctc ttcgatgtcc tcacatggct
     1561 gggatactgt aatagcacca tgaaccctat catctatccc ctcttcatgc gggacttcaa
     1621 gagggccctg ggcaggttcg tgccgtgtgt ccactgtccc ccggagcacc gggccagccc
     1681 cgcctccccc tccatgtgga cctctcacag tggtgccagg ccaggcctca gcctgcagca
     1741 ggtgctgccc ctgcctctgc cacccaactc agattcagac tcagcttcag ggggcacctc
     1801 gggcctgcag ctcacagccc agcttttgct gcctggagag gcgacccggg accccccgcc
     1861 acccaccagg gcccctactg tggtcaactt cttcgtcaca gactctgtgg agcctgagat
     1921 acggcagcat ccacttggtt cccccatgaa cagtgctggt ggtagcgcag gaggaccggt
     1981 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2041 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2101 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2161 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
     2221 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2281 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2341 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2401 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     2461 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     2521 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2581 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     2641 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2701 caagtccgga ctcagatctc gagctcaagc ttcgaattcg atggagctag aggtgccgga
     2761 cgaggcggag agcgccgaag cgggggccgt gacggcggaa gcggcctggt ctgcggagag
     2821 tggggcggcg gcaggtatga cccagaagaa ggctggcctc gcagaggccc ccttggtgac
     2881 agggcagcca ggccctggcc atgggaagaa gctgggccac cgaggcgtgg atgcgtcggg
     2941 agagactacg tataagaaga ccacctcgtc caccctgaag ggcgccatcc agctggggat
     3001 cgggtacacg gtgggcaacc tgagctctaa gccggagcgg gacgtgctca tgcaggactt
     3061 ctacgtgatg gagagcatct tcttccccag tgaagggagc aacttcaccc ctgcccacca
     3121 cttccaggat ttccgcttca agacctatgc acctgttgcc ttccgctact tccgggagct
     3181 ctttggcatc cgtccagatg actatttgta ctccctgtgc aatgagccac tcatcgagct
     3241 ctccaacccc ggggccagcg gctctgtctt ctacgtcacc agcgacgacg agttcatcat
     3301 caagactgtc atgcacaagg aggcagagtt cctgcagaag ctgctgcctg gctactacat
     3361 gaatctcaac cagaacccac ggacgctgct gcccaagttc tatgggctgt actgcgtgca
     3421 gtctggtggc aagaacatcc gcgtggtggt catgaacaat gtgctgcccc gtgtcgtcaa
     3481 aatgcacctt aagttcgacc tcaagggctc cacgtacaag cgcagggccg ccaagaagga
     3541 gaaggagaag agcctgccca cctacaagga cctggacttc atgcaggaca tgcccgaggg
     3601 gctgctgctg gactccgaca cctttggcgc cctggtcaag acgctgcagc gagactgcct
     3661 ggtgctggag agcttcaaga taatggacta cagcctgctg ctgggcgtgc acaacatcga
     3721 tcagcaggag cgagagcgcc aggccgaggg cgcccagagc aaggcggatg agaagcggcc
     3781 cgtggcccag aaggctctgt attccacggc catggagtct atccagggcg gagctgcccg
     3841 tggggaggcc attgagacag atgacacgat gggtgggatt ccagcagtga atgggcgcgg
     3901 ggagcgactg ctcctccaca tcgggatcat tgatattctg cagtcctaca ggttcatcaa
     3961 gaagttagaa cacacctgga aggccctcgt ccatgatggg gacactgtct cagtccaccg
     4021 gcccagcttc tatgcagagc gcttcttcaa gttcatgagc agcacggtgt tcgaggagag
     4081 ttcctccctg aagtcctctc catccaagaa aggacgtggt gccctgctgg cggtcaaacc
     4141 cctggggccc actgctgcct tctcagccag ccagatcccc agcgagagag aggatgtgca
     4201 gtatgacctg cggggggccc gcagctaccc cacgcttgag gatgaaggcc ggcctgacct
     4261 cctgccctgc accccaccgt cctttgagga agccaccaca gcctccatcg ccaccaccct
     4321 gtcgtccacc tccctctcca tcccagagcg gtccccttca gatacatcgg agcagccccg
     4381 gtacaggcgg cgcacgcagt cttcaggcca ggatggccgg ccccaggagg agccccatgc
     4441 ggaagacctg cagaagataa ctgtgcaggt ggagccagtg tgcggtgtgg gggttgtccc
     4501 caaggaggag ggtgcaggag tggaggtccc cccatgtggg gcatcggctg cagcctctgt
     4561 ggaaatagac gctgccagcc aggcctcaga gcctgccagc caggcctcag atgaggagga
     4621 tgcaccctct acagacatct atttttaagg atccaccgga tctagataac tgatcataat
     4681 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     4921 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4981 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5041 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5101 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5161 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5221 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5281 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5341 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5401 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5461 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     5521 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5581 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5641 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5701 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5761 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5821 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     5881 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5941 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6001 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6061 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6121 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6181 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6241 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6301 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6361 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     6421 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6481 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6541 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6601 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6661 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6721 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6781 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6841 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     6901 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6961 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     7021 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     7081 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     7141 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7201 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7261 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7321 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7501 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7621 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7681 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7861 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7921 gccttttgct cacatgttct ttcctgcgtt atcccctgat t
//
