LOCUS       sequence-181430-        6127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI(3,4,5)P3 single molecule fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  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 6127)
  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_GRP1
FEATURES             Location/Qualifiers
     source          1..6127
                     /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    2855..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2892..2911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2946..2965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2983..3438)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3120..3141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3332..3351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3465..3569
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3571..3928
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3571..3762
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3594..3614)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3779..3914
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3963..4757
                     /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             3963..4757
                     /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(4017..4036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4627..4646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4945..4964)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4989..5036
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5365..5953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5854..5873
                     /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 agatctcgag gaccgtttaa gatcccagaa gacgacggaa atgacctgac acacacgttc
     2281 ttcaacccag accgagaagg ctggctgctg aagctggggg gtcgtgtgaa gacctggaaa
     2341 cggcgctggt tcatcctcac agataactgc ctctactact ttgagtacac cacggacaag
     2401 gagcccaggg gcatcatccc cctggagaac ctcagcatca gggaggtgga ggacccccgg
     2461 aagccgaact gctttgagct gtataacccc agtcacaaag gtcaagtcat caaggcctgc
     2521 aagacggagg ccgatggccg tgtggtggag ggcaaccacg ttgtgtaccg gatctctgcc
     2581 cccagcccgg aggaaaagga ggagtggatg aagtccatca aagcaagcat cagtagggac
     2641 ccgttctatg acatgttggc cacgaggaaa aggaggattg ccaataagaa ataggaattc
     2701 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     2761 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2821 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2941 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3001 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3061 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3121 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3181 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3241 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3301 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3361 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3421 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3541 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3601 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3901 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4141 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4741 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4801 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4861 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4921 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4981 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5041 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5101 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5161 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6061 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6121 agttccg
//
