LOCUS       sequence-318717-        5013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged PH-FAPP1. PI(4)P biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5013)
  AUTHORS   Balla A, Tuymetova G, Tsiomenko A, Varnai P, Balla T
  TITLE     A plasma membrane pool of phosphatidylinositol 4-phosphate is 
            generated by phosphatidylinositol 4-kinase type-III alpha: studies 
            with the PH domains of the oxysterol binding protein and FAPP1.
  JOURNAL   Mol Biol Cell. 2005 Mar;16(3):1282-95. doi: 10.1091/mbc.e04-07-0578.
            Epub 2005 Jan 5.
  PUBMED    15635101
REFERENCE   2  (bases 1 to 5013)
  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: 
            "10.1091/mbc.e04-07-0578"; journalName: "Mol Biol Cell"; date: 
            "2005-03"; volume: "16"; issue: "3"; pages: "1282-95"
COMMENT     Sequence Label: GFP-PH-FAPP1
FEATURES             Location/Qualifiers
     source          1..5013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        91..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        395..598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     549..569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      983..992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(989..991,992..994,995..1708)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(989..991,992..994,995..1705)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1034..1055)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1295..1314)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1642..1663
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1831..1952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1868..1887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1922..1941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1959..2414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2096..2117)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2308..2327
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2441..2545
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2547..2904
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2547..2738
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2570..2590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2755..2890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2939..3733
                     /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             2939..3733
                     /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(2993..3012)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3603..3622
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3921..3940)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3965..4012
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4341..4929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4830..4849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      601 tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttcga
      661 attcaccatg gagggggtgt tgtacaagtg gaccaactat ctcacaggct ggcagcctcg
      721 ttggtttgtt ttagataatg gaatcttatc ctactatgat tcacaagatg atgtttgcaa
      781 agggagcaaa ggaagcataa agatggcagt ttgtgaaatt aaagttcatt cagcagacaa
      841 cacaagaatg gaattaatca ttcctggaga gcagcatttc tacatgaagg cagtgaatgc
      901 agctgaaaga cagaggtggc tggtcgctct ggggagctcc aaagcatgtt tgactgatac
      961 aaggactaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1021 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1081 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1141 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1201 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1261 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1321 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1381 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1441 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1501 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1561 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     1621 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1681 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     1741 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1801 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1861 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1921 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     1981 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2041 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2101 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2161 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2221 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2281 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2341 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2401 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2461 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2521 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     2581 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2641 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2701 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2761 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2821 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2881 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     2941 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3001 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3061 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3121 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3181 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3241 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3301 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3361 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3421 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     3481 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3541 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3601 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3661 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3721 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3781 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3841 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3901 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3961 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4021 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4081 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4141 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4981 gctcacatgt tctttcctgc gttatcccct gat
//
