LOCUS       sequence-287398-        8154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable PI4KA kinase-dead catalytic fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8154)
  AUTHORS   Zewe JP, Miller AM, Sangappa S, Wills RC, Goulden BD, Hammond GRV
  TITLE     Probing the subcellular distribution of phosphatidylinositol reveals
            a surprising lack at the plasma membrane.
  JOURNAL   J Cell Biol. 2020 Mar 2;219(3). pii: 133808. doi: 
            10.1083/jcb.201906127.
  PUBMED    32211893
REFERENCE   2  (bases 1 to 8154)
  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: "J Cell 
            Biol."; date: "2020-03-2"; volume: "219(3). pii: 133808. doi"; 
            pages: " 10.1083/jcb.201906127"
COMMENT     Sequence Label: mCherry-FKBP-PI4KAC1001(D1957A)
FEATURES             Location/Qualifiers
     source          1..8154
                     /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             613..1320
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1381..1701
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    4942..5063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4979..4998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5033..5052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5070..5525)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5207..5228)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5419..5438
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5552..5656
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5658..6015
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5658..5849
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5681..5701)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5866..6001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6050..6844
                     /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             6050..6844
                     /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(6104..6123)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6714..6733
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7032..7051)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7076..7123
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7452..8040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7941..7960
                     /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 gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgaag cgcggccgcg ggagcaggag gagcagctcg agcggcgctg
     1381 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag
     1441 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
     1501 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
     1561 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
     1621 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
     1681 gtggagcttc taaaactgga aagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     1741 gctggtggta gtgctggtgg tccgcgagct caagcttcga attccatggc cactgagagc
     1801 atccttcact ttgctggcta caacaagcag aacacaactc ttggggcaac tcagctgagc
     1861 gagcgcccgg cctgtgtgaa gaaagactac tccaacttca tggcatccct gaatctgcgc
     1921 aaccgctacg cgggcgaggt gtatggaatg attcggttct caggcaccac aggccagatg
     1981 tctgacctga acaaaatgat ggtccaggat ctacattcag ctttagaccg cagtcatcct
     2041 cagcactaca cgcaggccat gttcaagctg accgcaatgc tcattagcag taaagattgt
     2101 gacccgcagc tccttcatca tctgtgctgg ggtcccctcc ggatgttcaa tgagcatggc
     2161 atggagacgg ccctggcctg ctgggagtgg ctgctggctg gcaaggatgg agtggaagtg
     2221 ccgttcatgc gggagatggc aggggcctgg cacatgacgg tggagcagaa atttggcctg
     2281 ttttctgctg agataaagga agcagacccc ctggctgcct cggaagcaag tcaacccaaa
     2341 ccctgtcccc ccgaagtgac cccccactac atctggatcg acttcctggt gcagcggttt
     2401 gagatcgcca agtactgcag ctctgaccaa gtggagatct tctccagcct gctgcagcgc
     2461 tccatgtccc tgaacatcgg cggggccaag gggagcatga accggcacgt ggcggccatc
     2521 gggccccgct tcaagctgct gaccctgggg ctgtccctcc tgcatgccga tgtggttcca
     2581 aatgcaacca tccgcaatgt gcttcgcgag aagatctact ccactgcctt tgactacttc
     2641 agctgtcccc caaagttccc tactcaagga gagaagcggc tgcgtgaaga cataagcatc
     2701 atgattaaat tttggaccgc catgttctca gataagaagt acctgaccgc cagccagctt
     2761 gttcccccag ataatcagga cacccggagc aacctggaca taactgtcgg ctctcggcaa
     2821 caagccaccc aaggctggat caacacatac cccctgtcca gcggcatgtc caccatctcc
     2881 aagaaatcag gcatgtctaa gaaaaccaac cggggctccc agctgcacaa atactacatg
     2941 aagcgcagga cgctgctgct gtccctgctg gccactgaga tcgagcgtct catcacatgg
     3001 tacaacccgc tgtcagcccc ggaactggaa ctagaccagg ccggagagaa cagcgtggcc
     3061 aactggagat ctaagtacat cagcctgagt gagaagcagt ggaaggacaa cgtgaacctc
     3121 gcctggagca tctctcccta cctagccgtg cagctgcctg ccaggtttaa gaacacagaa
     3181 gccattggga acgaagtgac ccgtctcgtt cggttggacc cgggagccgt tagtgatgtg
     3241 cctgaagcaa tcaagttcct ggtcacctgg cacaccatcg acgccgatgc tccagagctc
     3301 agccatgtgc tgtgctgggc gcccacggac ccacccacag gcctctccta cttctccagc
     3361 atgtacccgc cgcaccctct cacagcgcag tacggggtga aagtcctgcg gtccttccct
     3421 ccggacgcca tcctcttcta catcccccag attgtgcagg ccctcaggta cgacaagatg
     3481 ggctatgtgc gggagtatat tctgtgggca gcgtctaaat cccagcttct ggcacaccag
     3541 ttcatctgga acatgaagac taacatttat ctagatgaag agggccacca gaaagaccct
     3601 gacatcggcg acctcctgga tcagttggta gaggagatca ccggctcctt gtccggccca
     3661 gcgaaggact tttaccagcg ggagtttgat ttctttaaca agatcaccaa cgtgtcggct
     3721 atcatcaagc cctaccctaa aggcgacgag agaaagaagg cttgtctgtc ggccctgtct
     3781 gaagtgaagg tgcagccggg ctgctacctg cccagcaacc ctgaggccat tgtgctggat
     3841 atcgactaca agtctgggac cccgatgcag agtgctgcaa aagccccata tctggccaag
     3901 ttcaaggtga agcgatgtgg tgttagtgaa cttgaaaaag aaggtctgcg gtgccgctca
     3961 gactccgagg atgagtgcag cacgcaggag gccgacggcc agaagatctc ctggcaggca
     4021 gccatcttca aggtgggaga cgactgccgg caggacatgc tggccctgca gatcatcgac
     4081 ctcttcaaga acatcttcca gctggtcggc ctggacctct ttgtttttcc ctaccgcgtg
     4141 gtggccactg cccctgggtg cggggtgatc gagtgcatcc ccgactgcac ctcccgggac
     4201 cagctgggcc gccagacaga cttcggcatg tacgactact tcacacgcca gtacggggat
     4261 gagtccactc tggccttcca gcaggcccgc tacaacttca tccgaagcat ggccgcctac
     4321 agcctcctgc tgttcctgct gcagatcaag gccagacaca acggcaacat tatgctggac
     4381 aagaagggtc atatcatcca catcgacttt ggcttcatgt ttgaaagctc gccgggcgac
     4441 aatctcggct gggaacccga catcaagctg acggatgaga tggtgatgat catggggggc
     4501 aagatggagg ccacaccctt caagtggttc atggagatgt gtgtccgagg ctacctggct
     4561 gtgcggccct acatggacgc ggtcgtctcc ctggtcactc tcatgttgga cacgggcctg
     4621 ccctgttttc gcggccagac aatcaagctc ttgaagcaca ggtttagccc caacatgact
     4681 gagcgcgagg ctgcaaattt catcatgaag gtcatccaga gctgcttcct cagcaacagg
     4741 agccggacct acgacatgat ccagtactat cagaatgaca tcccctactg agtcgacggt
     4801 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     4861 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4921 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4981 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5041 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     5101 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5161 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5221 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5281 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5341 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5401 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5461 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5521 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5581 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5641 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     5701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5761 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5821 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5881 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5941 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6001 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6061 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6121 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6181 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6241 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6301 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6361 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6421 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6481 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6541 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     6601 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6661 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6721 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6781 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6841 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6901 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6961 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     7021 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     7081 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     7141 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     7201 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     7261 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     7321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8101 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
