LOCUS       sequence-219820-        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        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    4711..4832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4748..4767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4802..4821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4839..5294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4976..4997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5188..5207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5321..5425
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5427..5784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5427..5618
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5450..5470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5635..5770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5819..6613
                     /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             5819..6613
                     /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(5873..5892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6483..6502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6801..6820)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6845..6892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7221..7809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7710..7729
                     /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 attctatgga aattaaagag
     1801 gaaggggcat cagaagaagg gcagcacttt cttcctacag cccaggccaa tgatcccggg
     1861 gactgtcagt tcacaagtat ccagaagact ccaaatgaac cgcagttgga attcatcctt
     1921 gcatgcaagg atctcgtggc tcctgtccgt gatcgtaaac tgaatacact ggtgcagatc
     1981 tccgtaatcc accccgtgga gcagagtctg acaagatact ccagcaccga aattgtggag
     2041 ggaacaaggg acccactgtt tttgactggt gtcacattcc catctgagta tcccatctat
     2101 gaggagacca aaataaaact aacagtctat gatgtcaagg ataagtctca tgacaccgtt
     2161 cgaaccagtg tcctaccaga acataaggat cccccgccag aagttgggcg aagtttcttg
     2221 ggctatgcca gttttaaagt gggagagctg ctgaagtcaa aggagcaatt gctggtcctg
     2281 agcctgagaa cttcagatgg tggcaaagtg gttggcacca tagaagtcag tgtcgtgaag
     2341 atgggggaga ttgaggatgg ggaagccgac cacatcacca cagatgtaca gggacaaaag
     2401 tgtgccctgg tatgtgaatg tacagccccg gaaagtgtga gcggaaaaga taacttacct
     2461 tttttgaatt cagtgttaaa gaacccagta tgtaaattat atagatttcc cacatctgac
     2521 aataagtgga tgcgaattcg agagcagatg tcagagagca ttctttcctt tcatattcct
     2581 aaggaattga tttcccttca cattaaagaa gatttgtgca gaaaccagga gataaaagaa
     2641 cttggtgagc tttctccaca ttgggacaat ctgcgaaaaa atgtccttac tcactgtgat
     2701 caaatggtga atatgtacca agacattctg acagaactta gcaaggaaac agggtcctct
     2761 ttcaaatcaa gcagcagcaa aggagagaaa acattagaat ttgttccaat aaatctacat
     2821 ctgcaaagaa tgcaggtaca cagccctcac ttgaaagatg ctctctacga tgtcatcact
     2881 gtgggagccc cagctgccca ttttcaggga tttaagaatg gtggtcttcg gaagctactc
     2941 catagatttg aaacagaaag aagaaatacc ggataccagt ttatttacta ttcacctgaa
     3001 aacacagcca aagcaaagga agttctcagc aacatcaatc aactacaacc tcttatagca
     3061 acccatgcag acctactgct taattctgca agccagcatt ctccagacag cttgaagaat
     3121 tctttaaaga tgctttcaga aaaaacagag ctttttgtac atgccttcaa ggatcaactt
     3181 gtcaggagtg ctcttttagc actctacact gcaaggccag gaggcattct taagaagcca
     3241 ccctctccta agagcagcac agaggagagc agtccccaag accaaccccc agtgatgaga
     3301 gggcaggact ccataccaca tcattcagac tatgatgagg aagagtggga cagggtgtgg
     3361 gccaatgtgg ggaagagcct gaactgcatt attgctatgg tggacaaact gattgaaaga
     3421 gatggtggca gtgaaggcag tggcggcaac aatgatggag aaaaggaacc ttcattaaca
     3481 gatgccattc cctctcaccc aagagaggac tggtatgaac agttgtatcc cctcatcctt
     3541 accctgaagg actgcatggg agaagtggtg aaccgagcca agcagtccct gacatttgtg
     3601 ctccttcagg aacttgcgta cagcttgccc cagtgtctga tgctgacgct aagaagagac
     3661 atcgtcttca gccaagcact tgctggattg gtttgtggtt ttatcatcaa attacagaca
     3721 agtctgtatg acccaggctt cctacagcag cttcacacag tggggttgat agtacaatat
     3781 gaaggactgc taagtacata cagcgatgaa attggaatgc tagaggacat ggccgttggc
     3841 atttccgatt taaagaaagt tgcatttaaa ataattgaag ccaaatccaa tgatgtgttg
     3901 ccagttataa caggaagacg agaacattac gtggtagagg tcaagcttcc agccagaatg
     3961 tttgagtcac tacctctaca gattaaagaa ggacagttgc ttcatgtgta tccagtactt
     4021 tttaatgttg gaatcaatga acagcaaact ctggctgaaa ggtttggaga tgtctctttg
     4081 caagaaagta ttaatcagga aaacttcgaa cttctacaag aatattacaa gatatttatg
     4141 gaaaagatgc ctcctgatta tatttcacat tttcaggaac aaaatgattt aaaagcattg
     4201 ctagaaaatc tccttcaaaa tatccaatcc aaaaaaagaa agaatgtaga aattatgtgg
     4261 ctggctgcaa cgatttgccg caaactgaat ggtattcgtt tcacctgttg taaaagtgcc
     4321 aaagacagga catcgatgtc agtgacactt gaacaatgct caatcttgag agatgagcac
     4381 cagttacaca aggacttctt tatccgagcg ctggattgca tgagaagaga aggatgccgc
     4441 atagagaatg tactgaagaa tatcaaatgc agaaagtatg ctttcaacat gctacagctg
     4501 atggctttcc ccaagtacta cagacctcca gaggggactt atggaaaagc tgacaccgca
     4561 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4621 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4681 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4741 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4801 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4861 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4921 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4981 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5041 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5101 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5161 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5221 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5281 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5341 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5401 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5461 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5521 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5581 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5641 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5701 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5761 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5821 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5881 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5941 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     6001 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     6061 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     6121 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     6181 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6241 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6301 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6361 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6421 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6481 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6541 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6601 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6661 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6721 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6781 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6841 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6901 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6961 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     7021 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     7081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7921 cat
//
