LOCUS       sequence-224781-        7923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable PI(3,4)P2 4-phosphatase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7923)
  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 7923)
  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: mCherry-FKBP-INPP4B
FEATURES             Location/Qualifiers
     source          1..7923
                     /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..1260
                     /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(698..716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     966..985
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1321..1641
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    4651..4772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4688..4707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4742..4761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4779..5234)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4916..4937)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5128..5147
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5261..5365
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5367..5724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5367..5558
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5390..5410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5575..5710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5759..6553
                     /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             5759..6553
                     /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(5813..5832)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6423..6442
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6741..6760)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6785..6832
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7161..7749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7650..7669
                     /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 caagggcgag gaggataaca tggccatcat caaggagttc
      601 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      661 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      721 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      781 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      901 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      961 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1021 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1081 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1141 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1201 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1261 tccggactca gatctcgaag cgcggccgcg ggagcaggag gagcagctcg agcggcgctg
     1321 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag
     1381 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
     1441 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
     1501 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
     1561 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
     1621 gtggagcttc taaaactgga aagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     1681 gctggtggta gtgctggtgg tccgcgagct caagcttcga attctatgga aattaaagag
     1741 gaaggggcat cagaagaagg gcagcacttt cttcctacag cccaggccaa tgatcccggg
     1801 gactgtcagt tcacaagtat ccagaagact ccaaatgaac cgcagttgga attcatcctt
     1861 gcatgcaagg atctcgtggc tcctgtccgt gatcgtaaac tgaatacact ggtgcagatc
     1921 tccgtaatcc accccgtgga gcagagtctg acaagatact ccagcaccga aattgtggag
     1981 ggaacaaggg acccactgtt tttgactggt gtcacattcc catctgagta tcccatctat
     2041 gaggagacca aaataaaact aacagtctat gatgtcaagg ataagtctca tgacaccgtt
     2101 cgaaccagtg tcctaccaga acataaggat cccccgccag aagttgggcg aagtttcttg
     2161 ggctatgcca gttttaaagt gggagagctg ctgaagtcaa aggagcaatt gctggtcctg
     2221 agcctgagaa cttcagatgg tggcaaagtg gttggcacca tagaagtcag tgtcgtgaag
     2281 atgggggaga ttgaggatgg ggaagccgac cacatcacca cagatgtaca gggacaaaag
     2341 tgtgccctgg tatgtgaatg tacagccccg gaaagtgtga gcggaaaaga taacttacct
     2401 tttttgaatt cagtgttaaa gaacccagta tgtaaattat atagatttcc cacatctgac
     2461 aataagtgga tgcgaattcg agagcagatg tcagagagca ttctttcctt tcatattcct
     2521 aaggaattga tttcccttca cattaaagaa gatttgtgca gaaaccagga gataaaagaa
     2581 cttggtgagc tttctccaca ttgggacaat ctgcgaaaaa atgtccttac tcactgtgat
     2641 caaatggtga atatgtacca agacattctg acagaactta gcaaggaaac agggtcctct
     2701 ttcaaatcaa gcagcagcaa aggagagaaa acattagaat ttgttccaat aaatctacat
     2761 ctgcaaagaa tgcaggtaca cagccctcac ttgaaagatg ctctctacga tgtcatcact
     2821 gtgggagccc cagctgccca ttttcaggga tttaagaatg gtggtcttcg gaagctactc
     2881 catagatttg aaacagaaag aagaaatacc ggataccagt ttatttacta ttcacctgaa
     2941 aacacagcca aagcaaagga agttctcagc aacatcaatc aactacaacc tcttatagca
     3001 acccatgcag acctactgct taattctgca agccagcatt ctccagacag cttgaagaat
     3061 tctttaaaga tgctttcaga aaaaacagag ctttttgtac atgccttcaa ggatcaactt
     3121 gtcaggagtg ctcttttagc actctacact gcaaggccag gaggcattct taagaagcca
     3181 ccctctccta agagcagcac agaggagagc agtccccaag accaaccccc agtgatgaga
     3241 gggcaggact ccataccaca tcattcagac tatgatgagg aagagtggga cagggtgtgg
     3301 gccaatgtgg ggaagagcct gaactgcatt attgctatgg tggacaaact gattgaaaga
     3361 gatggtggca gtgaaggcag tggcggcaac aatgatggag aaaaggaacc ttcattaaca
     3421 gatgccattc cctctcaccc aagagaggac tggtatgaac agttgtatcc cctcatcctt
     3481 accctgaagg actgcatggg agaagtggtg aaccgagcca agcagtccct gacatttgtg
     3541 ctccttcagg aacttgcgta cagcttgccc cagtgtctga tgctgacgct aagaagagac
     3601 atcgtcttca gccaagcact tgctggattg gtttgtggtt ttatcatcaa attacagaca
     3661 agtctgtatg acccaggctt cctacagcag cttcacacag tggggttgat agtacaatat
     3721 gaaggactgc taagtacata cagcgatgaa attggaatgc tagaggacat ggccgttggc
     3781 atttccgatt taaagaaagt tgcatttaaa ataattgaag ccaaatccaa tgatgtgttg
     3841 ccagttataa caggaagacg agaacattac gtggtagagg tcaagcttcc agccagaatg
     3901 tttgagtcac tacctctaca gattaaagaa ggacagttgc ttcatgtgta tccagtactt
     3961 tttaatgttg gaatcaatga acagcaaact ctggctgaaa ggtttggaga tgtctctttg
     4021 caagaaagta ttaatcagga aaacttcgaa cttctacaag aatattacaa gatatttatg
     4081 gaaaagatgc ctcctgatta tatttcacat tttcaggaac aaaatgattt aaaagcattg
     4141 ctagaaaatc tccttcaaaa tatccaatcc aaaaaaagaa agaatgtaga aattatgtgg
     4201 ctggctgcaa cgatttgccg caaactgaat ggtattcgtt tcacctgttg taaaagtgcc
     4261 aaagacagga catcgatgtc agtgacactt gaacaatgct caatcttgag agatgagcac
     4321 cagttacaca aggacttctt tatccgagcg ctggattgca tgagaagaga aggatgccgc
     4381 atagagaatg tactgaagaa tatcaaatgc agaaagtatg ctttcaacat gctacagctg
     4441 atggctttcc ccaagtacta cagacctcca gaggggactt atggaaaagc tgacaccgca
     4501 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4561 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4621 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4681 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4741 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4801 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4861 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4921 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4981 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5041 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5101 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5161 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5221 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5281 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5341 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5401 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5461 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5521 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5581 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5641 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5701 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5761 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5821 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5881 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5941 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     6001 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     6061 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     6121 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6181 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6241 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6301 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6361 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6421 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6481 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6541 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6601 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6661 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6721 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6781 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6841 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6901 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6961 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     7021 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7081 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7141 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7201 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7261 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7321 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7381 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7441 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7501 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7561 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7621 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7681 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7741 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7801 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7861 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7921 ccg
//
