LOCUS       sequence-224772-        7530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inactive plasma membrane targetted PI(3,4)P2 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-INPP4BC842A-CAAX
FEATURES             Location/Qualifiers
     source          1..7530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1254
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     polyA_signal    4258..4379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4295..4314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4349..4368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4386..4841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4523..4544)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4735..4754
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4868..4972
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4974..5331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4974..5165
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4997..5017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5182..5317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5366..6160
                     /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             5366..6160
                     /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(5420..5439)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6030..6049
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6348..6367)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6392..6439
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6768..7356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7257..7276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
      601 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
      661 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
      721 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
      781 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
      841 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      901 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
      961 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     1021 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     1081 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     1141 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     1201 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattccgga
     1261 ctcagatctc gagctcaagc ttcgaattct atggaaatta aagaggaagg ggcatcagaa
     1321 gaagggcagc actttcttcc tacagcccag gccaatgatc ccggggactg tcagttcaca
     1381 agtatccaga agactccaaa tgaaccgcag ttggaattca tccttgcatg caaggatctc
     1441 gtggctcctg tccgtgatcg taaactgaat acactggtgc agatctccgt aatccacccc
     1501 gtggagcaga gtctgacaag atactccagc accgaaattg tggagggaac aagggaccca
     1561 ctgtttttga ctggtgtcac attcccatct gagtatccca tctatgagga gaccaaaata
     1621 aaactaacag tctatgatgt caaggataag tctcatgaca ccgttcgaac cagtgtccta
     1681 ccagaacata aggatccccc gccagaagtt gggcgaagtt tcttgggcta tgccagtttt
     1741 aaagtgggag agctgctgaa gtcaaaggag caattgctgg tcctgagcct gagaacttca
     1801 gatggtggca aagtggttgg caccatagaa gtcagtgtcg tgaagatggg ggagattgag
     1861 gatggggaag ccgaccacat caccacagat gtacagggac aaaagtgtgc cctggtatgt
     1921 gaatgtacag ccccggaaag tgtgagcgga aaagataact tacctttttt gaattcagtg
     1981 ttaaagaacc cagtatgtaa attatataga tttcccacat ctgacaataa gtggatgcga
     2041 attcgagagc agatgtcaga gagcattctt tcctttcata ttcctaagga attgatttcc
     2101 cttcacatta aagaagattt gtgcagaaac caggagataa aagaacttgg tgagctttct
     2161 ccacattggg acaatctgcg aaaaaatgtc cttactcact gtgatcaaat ggtgaatatg
     2221 taccaagaca ttctgacaga acttagcaag gaaacagggt cctctttcaa atcaagcagc
     2281 agcaaaggag agaaaacatt agaatttgtt ccaataaatc tacatctgca aagaatgcag
     2341 gtacacagcc ctcacttgaa agatgctctc tacgatgtca tcactgtggg agccccagct
     2401 gcccattttc agggatttaa gaatggtggt cttcggaagc tactccatag atttgaaaca
     2461 gaaagaagaa ataccggata ccagtttatt tactattcac ctgaaaacac agccaaagca
     2521 aaggaagttc tcagcaacat caatcaacta caacctctta tagcaaccca tgcagaccta
     2581 ctgcttaatt ctgcaagcca gcattctcca gacagcttga agaattcttt aaagatgctt
     2641 tcagaaaaaa cagagctttt tgtacatgcc ttcaaggatc aacttgtcag gagtgctctt
     2701 ttagcactct acactgcaag gccaggaggc attcttaaga agccaccctc tcctaagagc
     2761 agcacagagg agagcagtcc ccaagaccaa cccccagtga tgagagggca ggactccata
     2821 ccacatcatt cagactatga tgaggaagag tgggacaggg tgtgggccaa tgtggggaag
     2881 agcctgaact gcattattgc tatggtggac aaactgattg aaagagatgg tggcagtgaa
     2941 ggcagtggcg gcaacaatga tggagaaaag gaaccttcat taacagatgc cattccctct
     3001 cacccaagag aggactggta tgaacagttg tatcccctca tccttaccct gaaggactgc
     3061 atgggagaag tggtgaaccg agccaagcag tccctgacat ttgtgctcct tcaggaactt
     3121 gcgtacagct tgccccagtg tctgatgctg acgctaagaa gagacatcgt cttcagccaa
     3181 gcacttgctg gattggtttg tggttttatc atcaaattac agacaagtct gtatgaccca
     3241 ggcttcctac agcagcttca cacagtgggg ttgatagtac aatatgaagg actgctaagt
     3301 acatacagcg atgaaattgg aatgctagag gacatggccg ttggcatttc cgatttaaag
     3361 aaagttgcat ttaaaataat tgaagccaaa tccaatgatg tgttgccagt tataacagga
     3421 agacgagaac attacgtggt agaggtcaag cttccagcca gaatgtttga gtcactacct
     3481 ctacagatta aagaaggaca gttgcttcat gtgtatccag tactttttaa tgttggaatc
     3541 aatgaacagc aaactctggc tgaaaggttt ggagatgtct ctttgcaaga aagtattaat
     3601 caggaaaact tcgaacttct acaagaatat tacaagatat ttatggaaaa gatgcctcct
     3661 gattatattt cacattttca ggaacaaaat gatttaaaag cattgctaga aaatctcctt
     3721 caaaatatcc aatccaaaaa aagaaagaat gtagaaatta tgtggctggc tgcaacgatt
     3781 tgccgcaaac tgaatggtat tcgtttcacc tgtgctaaaa gtgccaaaga caggacatcg
     3841 atgtcagtga cacttgaaca atgctcaatc ttgagagatg agcaccagtt acacaaggac
     3901 ttctttatcc gagcgctgga ttgcatgaga agagaaggat gccgcataga gaatgtactg
     3961 aagaatatca aatgcagaaa gtatgctttc aacatgctac agctgatggc tttccccaag
     4021 tactacagac ctccagaggg gacttatgga aaagctgaca ccgcagtcga caaccctcct
     4081 gatgagagtg gccccggctg catgagctgc aagtgtgtgc tctcctgagg atccaccgga
     4141 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4201 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4261 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4321 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     4381 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4441 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4501 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4561 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4621 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4681 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4741 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4801 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4861 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4921 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4981 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5101 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5161 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5221 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5281 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     5341 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5401 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5461 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5521 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     5581 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5641 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5701 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5761 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5821 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5881 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5941 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6001 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6061 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6121 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6181 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6241 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6301 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     6361 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     6421 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6481 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     6541 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6601 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6661 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6721 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6781 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6841 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6901 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6961 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7021 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7081 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7141 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7201 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7261 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7321 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7381 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7441 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     7501 attagttcat agcccatata tggagttccg
//
