LOCUS       sequence-181413-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI(4,5)P2 single molecule fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  AUTHORS   Ji C, Zhang Y, Xu P, Xu T, Lou X
  TITLE     Nanoscale Landscape of Phosphoinositides Revealed by Specific 
            Pleckstrin Homology (PH) Domains Using Single-molecule 
            Superresolution Imaging in the Plasma Membrane.
  JOURNAL   J Biol Chem. 2015 Nov 6;290(45):26978-93. doi: 
            10.1074/jbc.M115.663013. Epub 2015 Sep 22.
  PUBMED    26396197
REFERENCE   2  (bases 1 to 6145)
  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: 
            "10.1074/jbc.M115.663013"; journalName: "J Biol Chem"; date: 
            "2015-11-6- 6"; volume: "290"; issue: "45"; pages: "26978-93"
COMMENT     Sequence Label: iRFP-PAmCherry1-PH_PLCd1
FEATURES             Location/Qualifiers
     source          1..6145
                     /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      544..553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             550..1497
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     regulatory      1507..1516
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1513..2220
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1658..1676)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2873..2994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2910..2929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2964..2983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3001..3456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3138..3159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3350..3369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3483..3587
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3589..3946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3589..3780
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3612..3632)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3797..3932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3981..4775
                     /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             3981..4775
                     /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(4035..4054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4645..4664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4963..4982)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5007..5054
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5383..5971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5872..5891
                     /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 gctagcggta
      541 cccgccacca tggctgaagg atccgtcgcc aggcagcctg acctcttgac ctgcgacgat
      601 gagccgatcc atatccccgg tgccatccaa ccgcatggac tgctgctcgc cctcgccgcc
      661 gacatgacga tcgttgccgg cagcgacaac cttcccgaac tcaccggact ggcgatcggc
      721 gccctgatcg gccgctctgc ggccgatgtc ttcgactcgg agacgcacaa ccgtctgacg
      781 atcgccttgg ccgagcccgg ggcggccgtc ggagcaccga tcactgtcgg cttcacgatg
      841 cgaaaggacg caggcttcat cggctcctgg catcgccatg atcagctcat cttcctcgag
      901 ctcgagcctc cccagcggga cgtcgccgag ccgcaggcgt tcttccgccg caccaacagc
      961 gccatccgcc gcctgcaggc cgccgaaacc ttggaaagcg cctgcgccgc cgcggcgcaa
     1021 gaggtgcgga agattaccgg cttcgatcgg gtgatgatct atcgcttcgc ctccgacttc
     1081 agcggcgaag tgatcgcaga ggatcggtgc gccgaggtcg agtcaaaact aggcctgcac
     1141 tatcctgcct caaccgtgcc ggcgcaggcc cgtcggctct ataccatcaa cccggtacgg
     1201 atcattcccg atatcaatta tcggccggtg ccggtcaccc cagacctcaa tccggtcacc
     1261 gggcggccga ttgatcttag cttcgccatc ctgcgcagcg tctcgcccgt ccatctggaa
     1321 ttcatgcgca acataggcat gcacggcacg atgtcgatct cgattttgcg cggcgagcga
     1381 ctgtggggat tgatcgtttg ccatcaccga acgccgtact acgtcgatct cgatggccgc
     1441 caagcctgcg agctagtcgc ccaggttctg gcctggcaga tcggcgtgat ggaagagcca
     1501 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat taaggagttc
     1561 atgcgcttca aggtgcacat ggaggggtcc gtgaacggcc acgtgttcga gatcgagggc
     1621 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     1681 ggccccctgc ccttcacctg ggacatcctg agccctcagt tcatgtacgg ctccaatgcc
     1741 tacgtgaagc accccgccga catccccgac tactttaagc tgtccttccc cgagggcttc
     1801 aagtgggagc gcgtgatgaa attcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1861 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1921 gacggccccg tgatgcagaa gaagaccatg ggctgggagg ccctctccga gcggatgtac
     1981 cccgaggacg gcgccctgaa gggcgaggtc aagcccagag tgaagctgaa ggacggcggc
     2041 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2101 gcctacaacg tcaaccgcaa gctggacatc accagccaca acgaggacta caccatcgtg
     2161 gagcagtacg agagagccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     2221 agatctcccg ggggcatgga ctcgggccgg gacttcctga ccctgcacgg cctacaggat
     2281 gatgaggatc tacaggcgct gctgaagggc agccagctcc tgaaggtgaa gtccagctca
     2341 tggaggagag agcgcttcta caagttgcag gaggactgca agaccatctg gcaggagtcc
     2401 cgcaaggtca tgcggacccc ggagtcccag ctgttctcca tcgaggacat tcaggaggtg
     2461 cgaatggggc accgcacgga gggtctggag aagttcgccc gtgatgtgcc cgaggaccgc
     2521 tgcttctcca ttgtcttcaa ggaccagcgc aatacactag acctcatcgc cccatcgcca
     2581 gctgatgccc agcactgggt gctggggctg cacaagatca tccaccactc aggctccatg
     2641 gaccagcgtc agaagctaca gcactggatt cactcctgct tgcgaaaagc tgacaaaaac
     2701 aaggacaaca agatgagctt caaggagctg cagaacttcc tgaaggagta agcggccgcg
     2761 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2821 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2881 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2941 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3001 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3061 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3541 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3601 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3721 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3781 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3841 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3901 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3961 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4021 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4081 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4141 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4201 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4261 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4321 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4381 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4441 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4501 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4561 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4621 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4681 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4741 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4801 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4861 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4921 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4981 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5041 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5101 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5161 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5221 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5281 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5341 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5401 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5461 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5521 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5581 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5641 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5701 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5761 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5821 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5881 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5941 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6001 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6061 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     6121 ttcatagccc atatatggag ttccg
//
