LOCUS       sequence-473813-        7727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endosome-targeted inositol polyphosphate 5 phosphatase (INPP5E); 
            hydrolyzes 5-phosphate in PI(3,4,5)P3 and PI(4,5)P2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7727)
  AUTHORS   Sahan AZ, Chen M, Su Q, Li Q, Wang D, Zhang J
  TITLE     Lysosomal PIP(3) revealed by genetically encoded lipid biosensors.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Apr;122(13):e2426929122. doi: 
            10.1073/pnas.2426929122. Epub 2025 Mar 24.
  PUBMED    40127277
REFERENCE   2  (bases 1 to 7727)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-04"; volume: "122(13):e2426929122. 
            doi"; pages: " 10.1073/pnas.2426929122. Epub 2025 Mar 24"
COMMENT     Sequence Label: pcDNA3-Endo-INPP5E
FEATURES             Location/Qualifiers
     source          1..7727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      902..911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             923..940
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             944..976
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(980..988,989..1003)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2315..3022
                     /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(2460..2478)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2728..2747
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3281..3299)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3282..3299)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3319..3336)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3325..3549
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3595..4023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3682..3701)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3892..3913
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4032..4052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4037..4366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4217..4352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4279..4298
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4433..5227
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4487..4506)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5097..5116
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5401..5522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5438..5457)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5492..5511
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5571..5587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5571..5587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5584..5606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5595..5611
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5619..5625,5626..5643,5644..5649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5664..5685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5802..5819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5973..6561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6053..6072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6732..7523,7524..7592))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7355..7374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7593..7697)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agcttgcggc
      901 cgccaccatg gtgcggggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg
      961 tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccct cggatcttgc
     1021 agactacaag ctccgcgcgc agccgctcct ggtgcgggcc cacagcagcc tgggccccgg
     1081 ccggccgcgg agccccctgg cctgcgacga ctgttccctt cgctcggcca aatcctcctt
     1141 cagcctcctg gcgcccatcc gcagcaagga cgtccgcagc aggagttacc tggagggcag
     1201 cctcctggcc agcggggccc tgttgggggc ggatgagctg gcccgctact tcccagaccg
     1261 gaacgtggca ctcttcgtgg ccacctggaa catgcagggc cagaaggagc tcccgcccag
     1321 cctggacgag ttcctgctcc cagccgaggc cgactatgcc caggacctgt atgtcatcgg
     1381 ggtccaggag ggctgttctg acaggcggga gtgggagact cgtctgcagg agacgctggg
     1441 cccgcactat gtgctgctgt cctcggcggc ccacggcgtg ctctacatgt cgctcttcat
     1501 ccgcagggac ctcatctggt tctgctcaga ggtggagtgc tccacggtga ccacacgcat
     1561 cgtgtctcag atcaagacca agggggcctt gggcatcagc ttcacctttt ttggcacttc
     1621 cttcctcttc atcacgtccc acttcacctc aggtgacggg aaggtggcgg agcggctgct
     1681 ggactacacc aggactgtac aagccctggt cctgcccaga aatgtgcccg acaccaaccc
     1741 ctatcgctcc agcgcagcgg acgtcaccac ccgcttcgat gaggtgttct ggtttggaga
     1801 cttcaacttc cgcctgagtg gcgggcgcac agtcgtggac gccctcctgt gccagggcct
     1861 ggtggtggac gtgccggcgc tgctgcagca cgaccagctc atccgggaga tgcggaaagg
     1921 gtccatcttc aagggcttcc aggagccgga catccacttc ctcccatcat acaagtttga
     1981 catcgggaag gacacgtacg acagcacctc caagcagagg acgccctcat acacggaccg
     2041 cgtcttgtac agaagccgcc acaagggtga catctgtcct gtgagctact cttcctgccc
     2101 cgggatcaag acgtccgacc accgccctgt gtatggcctc ttccgggtga aagtgaggcc
     2161 ggggcgagac aacattccgt tggcagctgg caaatttgat agagaactgt acttactagg
     2221 aattaaaaga cggatttcga aggagattca gaggcagcaa gcactacaga gtcagaactc
     2281 cagcaccatc gcctccgttt ctgctagcga tcccatggtg agcaagggcg aggaggataa
     2341 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     2401 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2461 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2521 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2581 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2641 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2701 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2761 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2821 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     2881 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     2941 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     3001 cggcatggac gagctgtaca agaaggaatt cgaaagtgat gccatgttcg ctgctgaaag
     3061 agcccctgac tgggtggatg ctgaggaatg ccatcggtgc agagtacagt ttggggtggt
     3121 gacccgcaag catcactgcc gagcatgtgg gcagatcttc tgtggcaagt gctcctccaa
     3181 gtactccacc atccccaagt tcggcattga gaaggaggtg cgcgtgtgtg agccctgcta
     3241 tgagcagctg aacaagaagg cataatctag agggccctat tctatagtgt cacctaaatg
     3301 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3361 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3421 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3481 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3541 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3601 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3661 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3721 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3781 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3841 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3901 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3961 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4021 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4081 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4141 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4201 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4261 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4321 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4381 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4441 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4501 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4561 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4621 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4681 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4741 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4801 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4861 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4921 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4981 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5041 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5101 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5161 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5221 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5281 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5341 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5401 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5461 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5521 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5581 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5641 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5701 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5761 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5821 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5881 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5941 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6001 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6061 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6661 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6721 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6781 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6841 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6901 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6961 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7021 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7081 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7141 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7201 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7261 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7321 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7381 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7441 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7501 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7561 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7621 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7681 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgt
//
