LOCUS       sequence-224762-        7530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasma membrane-targetted PI(3,4)P2 4-phosphatase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7530)
  AUTHORS   Goulden BD, Pacheco J, Dull A, Zewe JP, Deiters A, Hammond GRV
  TITLE     A high-avidity biosensor reveals plasma membrane PI(3,4)P2 is 
            predominantly a class I PI3K signaling product.
  JOURNAL   J Cell Biol. 2019 Mar 4;218(3):1066-1079. doi: 
            10.1083/jcb.201809026. Epub 2018 Dec 27.
  PUBMED    30591513
REFERENCE   2  (bases 1 to 7530)
  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"; doi: 
            "10.1083/jcb.201809026"; journalName: "J Cell Biol"; date: 
            "2019-03-4- 4"; volume: "218"; issue: "3"; pages: "1066-1079"
COMMENT     Sequence Label: TagBFP2-INPP4B-CAAX
FEATURES             Location/Qualifiers
     source          1..7530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             249..950
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     polyA_signal    3954..4075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3991..4010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4045..4064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4082..4537)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4219..4240)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4431..4450
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4564..4668
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4670..5027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4670..4861
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4693..4713)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4878..5013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5062..5856
                     /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             5062..5856
                     /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(5116..5135)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5726..5745
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6044..6063)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6088..6135
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6464..7052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6953..6972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7227..7530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
      301 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
      361 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
      421 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
      481 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
      541 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
      601 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
      661 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
      721 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac gccaagacca
      781 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
      841 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
      901 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat tccggactca
      961 gatctcgagc tcaagcttcg aattctatgg aaattaaaga ggaaggggca tcagaagaag
     1021 ggcagcactt tcttcctaca gcccaggcca atgatcccgg ggactgtcag ttcacaagta
     1081 tccagaagac tccaaatgaa ccgcagttgg aattcatcct tgcatgcaag gatctcgtgg
     1141 ctcctgtccg tgatcgtaaa ctgaatacac tggtgcagat ctccgtaatc caccccgtgg
     1201 agcagagtct gacaagatac tccagcaccg aaattgtgga gggaacaagg gacccactgt
     1261 ttttgactgg tgtcacattc ccatctgagt atcccatcta tgaggagacc aaaataaaac
     1321 taacagtcta tgatgtcaag gataagtctc atgacaccgt tcgaaccagt gtcctaccag
     1381 aacataagga tcccccgcca gaagttgggc gaagtttctt gggctatgcc agttttaaag
     1441 tgggagagct gctgaagtca aaggagcaat tgctggtcct gagcctgaga acttcagatg
     1501 gtggcaaagt ggttggcacc atagaagtca gtgtcgtgaa gatgggggag attgaggatg
     1561 gggaagccga ccacatcacc acagatgtac agggacaaaa gtgtgccctg gtatgtgaat
     1621 gtacagcccc ggaaagtgtg agcggaaaag ataacttacc ttttttgaat tcagtgttaa
     1681 agaacccagt atgtaaatta tatagatttc ccacatctga caataagtgg atgcgaattc
     1741 gagagcagat gtcagagagc attctttcct ttcatattcc taaggaattg atttcccttc
     1801 acattaaaga agatttgtgc agaaaccagg agataaaaga acttggtgag ctttctccac
     1861 attgggacaa tctgcgaaaa aatgtcctta ctcactgtga tcaaatggtg aatatgtacc
     1921 aagacattct gacagaactt agcaaggaaa cagggtcctc tttcaaatca agcagcagca
     1981 aaggagagaa aacattagaa tttgttccaa taaatctaca tctgcaaaga atgcaggtac
     2041 acagccctca cttgaaagat gctctctacg atgtcatcac tgtgggagcc ccagctgccc
     2101 attttcaggg atttaagaat ggtggtcttc ggaagctact ccatagattt gaaacagaaa
     2161 gaagaaatac cggataccag tttatttact attcacctga aaacacagcc aaagcaaagg
     2221 aagttctcag caacatcaat caactacaac ctcttatagc aacccatgca gacctactgc
     2281 ttaattctgc aagccagcat tctccagaca gcttgaagaa ttctttaaag atgctttcag
     2341 aaaaaacaga gctttttgta catgccttca aggatcaact tgtcaggagt gctcttttag
     2401 cactctacac tgcaaggcca ggaggcattc ttaagaagcc accctctcct aagagcagca
     2461 cagaggagag cagtccccaa gaccaacccc cagtgatgag agggcaggac tccataccac
     2521 atcattcaga ctatgatgag gaagagtggg acagggtgtg ggccaatgtg gggaagagcc
     2581 tgaactgcat tattgctatg gtggacaaac tgattgaaag agatggtggc agtgaaggca
     2641 gtggcggcaa caatgatgga gaaaaggaac cttcattaac agatgccatt ccctctcacc
     2701 caagagagga ctggtatgaa cagttgtatc ccctcatcct taccctgaag gactgcatgg
     2761 gagaagtggt gaaccgagcc aagcagtccc tgacatttgt gctccttcag gaacttgcgt
     2821 acagcttgcc ccagtgtctg atgctgacgc taagaagaga catcgtcttc agccaagcac
     2881 ttgctggatt ggtttgtggt tttatcatca aattacagac aagtctgtat gacccaggct
     2941 tcctacagca gcttcacaca gtggggttga tagtacaata tgaaggactg ctaagtacat
     3001 acagcgatga aattggaatg ctagaggaca tggccgttgg catttccgat ttaaagaaag
     3061 ttgcatttaa aataattgaa gccaaatcca atgatgtgtt gccagttata acaggaagac
     3121 gagaacatta cgtggtagag gtcaagcttc cagccagaat gtttgagtca ctacctctac
     3181 agattaaaga aggacagttg cttcatgtgt atccagtact ttttaatgtt ggaatcaatg
     3241 aacagcaaac tctggctgaa aggtttggag atgtctcttt gcaagaaagt attaatcagg
     3301 aaaacttcga acttctacaa gaatattaca agatatttat ggaaaagatg cctcctgatt
     3361 atatttcaca ttttcaggaa caaaatgatt taaaagcatt gctagaaaat ctccttcaaa
     3421 atatccaatc caaaaaaaga aagaatgtag aaattatgtg gctggctgca acgatttgcc
     3481 gcaaactgaa tggtattcgt ttcacctgtt gtaaaagtgc caaagacagg acatcgatgt
     3541 cagtgacact tgaacaatgc tcaatcttga gagatgagca ccagttacac aaggacttct
     3601 ttatccgagc gctggattgc atgagaagag aaggatgccg catagagaat gtactgaaga
     3661 atatcaaatg cagaaagtat gctttcaaca tgctacagct gatggctttc cccaagtact
     3721 acagacctcc agaggggact tatggaaaag ctgacaccgc agtcgacaac cctcctgatg
     3781 agagtggccc cggctgcatg agctgcaagt gtgtgctctc ctgaggatcc accggatcta
     3841 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3901 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3961 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4021 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     4081 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4141 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4201 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4261 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4321 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4381 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4441 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4501 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     4561 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4621 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     4681 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4741 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4801 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4861 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4921 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4981 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     5041 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5101 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5161 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5221 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5281 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5341 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5401 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5461 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5521 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5581 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5641 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5701 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5761 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5821 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5881 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5941 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6001 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     6061 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     6121 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     6181 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     6241 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     6301 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6601 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7081 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7141 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
     7201 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     7261 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     7321 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7381 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     7441 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     7501 atctacgtat tagtcatcgc tattaccatg
//
