LOCUS       sequence-269440-        7557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable PI4KB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7557)
  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 7557)
  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-PI4KB
FEATURES             Location/Qualifiers
     source          1..7557
                     /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    4345..4466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4382..4401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4436..4455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4473..4928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4610..4631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4822..4841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4955..5059
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5061..5418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5061..5252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5084..5104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5269..5404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5453..6247
                     /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             5453..6247
                     /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(5507..5526)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6117..6136
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6435..6454)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6479..6526
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6855..7443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7344..7363
                     /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 attctgcagt cgacatggga
     1801 gatacagtag tggagcctgc ccccttgaag ccaacttctg agcccacttc tggcccacca
     1861 gggaataatg gggggtccct gctaagtgtc atcacggagg gggtcgggga actatcagtg
     1921 attgaccctg aggtggccca gaaggcctgc caggaggtgt tggagaaagt caagcttttg
     1981 catggaggcg tggcagtctc tagcagaggc accccactgg agttggtcaa tggggatggt
     2041 gtggacagtg agatccgttg cctagatgat ccacctgccc agatcaggga ggaggaagat
     2101 gagatggggg ccgctgtggc ctcaggcaca gccaaaggag caagaagacg gcggcagaac
     2161 aactcagcta aacagtcttg gctgctgagg ctgtttgagt caaaactgtt tgacatctcc
     2221 atggccattt catacctgta taactccaag gagcctggag tacaagccta cattggcaac
     2281 cggctcttct gctttcgcaa cgaggacgtg gacttctatc tgccccagtt gcttaacatg
     2341 tacatccaca tggatgagga cgtgggtgat gccattaagc cctacatagt ccaccgttgc
     2401 cgccagagca ttaacttttc cctccagtgt gccctgttgc ttggggccta ttcttcagac
     2461 atgcacattt ccactcaacg acactcccgt gggaccaagc tacggaagct gatcctctca
     2521 gatgagctaa agccagctca caggaagagg gagctgccct ccttgagccc ggcccctgac
     2581 acagggctgt ctccctccaa aaggactcac cagcgctcta agtcagatgc cactgccagc
     2641 ataagtctca gcagcaacct gaaacgaaca gccagcaacc ctaaagtgga gaatgaggat
     2701 gagcctgttc gactggctcc tgagagagaa ttcatcaagt ccctgatggc gatcggcaag
     2761 cggctggcca cgctccccac caaagagcag aaaacacaga ggctgatctc agagctctcc
     2821 ctgctcaacc ataagctccc tgcccgagtc tggctgccca ctgctggctt tgaccaccac
     2881 gtggtccgtg taccccacac acaggctgtt gtcctcaact ccaaggacaa ggctccctac
     2941 ctgatttatg tggaagtcct tgaatgtgaa aactttgaca ccaccagtgt ccctgcccgg
     3001 atccccgaga accgaattcg gagtacgagg tccgtagaaa acttgcccga atgtggtatt
     3061 acccatgagc agcgagctgg cagcttcagc actgtgccca actatgacaa cgatgatgag
     3121 gcctggtcgg tggatgacat aggcgagctg caagtggagc tccccgaagt gcataccaac
     3181 agctgtgaca acatctccca gttctctgtg gacagcatca ccagccagga gagcaaggag
     3241 cctgtgttca ttgcagcagg ggacatccgc cggcgccttt cggaacagct ggctcatacc
     3301 ccgacagcct tcaaacgaga cccagaagat ccttctgcag ttgctctcaa agagccctgg
     3361 caggagaaag tacggcggat cagagagggc tccccctacg gccatctccc caattggcgg
     3421 ctcctgtcag tcattgtcaa gtgtggggat gaccttcggc aagagcttct ggcctttcag
     3481 gtgttgaagc aactgcagtc catttgggaa caggagcgag tgcccctttg gatcaagcca
     3541 tacaagattc ttgtgatttc ggctgatagt ggcatgattg aaccagtggt caatgctgtg
     3601 tccatccatc aggtgaagaa acagtcacag ctctccttgc tcgattactt cctacaggag
     3661 cacggcagtt acaccactga ggcattcctc agtgcacagc gcaattttgt gcaaagttgt
     3721 gctgggtact gcttggtctg ctacctgctg caagtcaagg acagacacaa tgggaatatc
     3781 cttttggacg cagaaggcca catcatccac atcgactttg gcttcatcct ctccagctca
     3841 ccccgaaatc tgggctttga gacgtcagcc tttaagctga ccacagagtt tgtggatgtg
     3901 atgggcggcc tggatggcga catgttcaac tactataaga tgctgatgct gcaagggctg
     3961 attgccgctc ggaaacacat ggacaaggtg gtgcagatcg tggagatcat gcagcaaggt
     4021 tctcagcttc cttgcttcca tggctccagc accattcgaa acctcaaaga gaggttccac
     4081 atgagcatga ctgaggagca gctgcagctg ctggtggagc agatggtgga tggcagtatg
     4141 cggtctatca ccaccaaact ctatgacggc ttccagtacc tcaccaacgg catcatgtga
     4201 ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag ccataccaca
     4261 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4321 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4381 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4441 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     4501 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4561 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4621 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4681 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4741 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4801 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4861 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4921 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4981 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5041 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5101 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5161 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5221 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5281 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5341 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5401 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5461 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5521 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5581 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5641 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5701 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5761 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5821 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5881 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5941 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     6001 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6061 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6121 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6181 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6241 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6301 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6361 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6421 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6481 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6541 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6601 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6661 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6961 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     7021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7501 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
