LOCUS       sequence-364316-        7421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded single-color sensor for monitoring plasma 
            membrane PI(3,4,5)P2 dynamics in living cells. Based on 
            dimerization-dependent red fluorescent protein (ddRFP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7421)
  AUTHORS   Hertel F, Li S, Chen M, Pott L, Mehta S, Zhang J
  TITLE     Fluorescent Biosensors for Multiplexed Imaging of Phosphoinositide 
            Dynamics.
  JOURNAL   ACS Chem Biol. 2020 Jan 17;15(1):33-38. doi: 
            10.1021/acschembio.9b00691. Epub 2020 Jan 6.
  PUBMED    31855412
REFERENCE   2  (bases 1 to 7421)
  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: 
            "10.1021/acschembio.9b00691"; journalName: "ACS Chem Biol"; date: 
            "2020-01-17- 17"; volume: "15"; issue: "1"; pages: "33-38"
COMMENT     Sequence Label: pcDNA3-Lyn-dInPAkt
FEATURES             Location/Qualifiers
     source          1..7421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      946..955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2974..2992)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2975..2992)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3012..3029)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3018..3242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3288..3716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3375..3394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3585..3606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3725..3745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3730..4059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3910..4045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3972..3991
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4126..4920
                     /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(4180..4199)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4790..4809
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5094..5215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5131..5150)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5185..5204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5264..5280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5264..5280)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5277..5299)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5288..5304
                     /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(5312..5318,5319..5336,5337..5342))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5357..5378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5495..5512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5666..6254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5746..5765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6425..7216,7217..7285))
                     /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     7048..7067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7286..7390)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gccaccatgg gctgcatcaa gagcaagcgc aaggacaagg atccggccac catggtgagc
      961 aagagcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcttgga gggctccatg
     1021 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1081 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     1141 ccccagatca tgtacggctc caaggcgtac gtgaagcacc ccgccgacgt ccccgattac
     1201 atgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgcactt cgaggacggc
     1261 ggtctggtga ccgtgactca ggacacctcc ctgcaggacg gcacgctgat ctacaaggtg
     1321 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccttgggc
     1381 tgggattatg ccaccgagcg cctgtacccc gaagacggcg tgctgaaggg cgagcttctg
     1441 gggcgcctga agctgaagga cggcggcctc aacctggtgg agttcaagac catctacatg
     1501 gccaagaagc ccgtgcaact gcccggctac tacttcgtgg acaccaagct gggcatcacc
     1561 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     1621 ctggggcatg gcaccggcag cgcatgcatg aacgacgtag ccattgtgaa ggagggctgg
     1681 ctgcacaaac gaggggaata tattaaaacc tggcggccac gctacttcct cctcaagaat
     1741 gatggcacct ttattggcta caaggaacgg cctcaggatg tggagcagcg cgagtcccca
     1801 ctcaacaact tctcagtggc acaatgtcag ctgatgaaga cagagcggcc gaggcccaac
     1861 accttcatca tccgctgcct gcagtggacc acggtcattg agcgcacctt ccatgtggaa
     1921 acgcctgagg agcgggaaga gtggaccacc gccattcaga ctgtggctga tggactcaaa
     1981 cggcaggagg aggagacgat ggacttccgg tcaggttcac ccagtgacaa ctcaggtgct
     2041 gaggagatgg aggtggccct ggccaagccc aagcaccgtg tgaccatgaa cgagtttgag
     2101 tacctgaagc tactgggcaa gggcaccttt gggaaggtgg tcgacggcgg tagcgtggca
     2161 gaagaggaag atgacgagga agaggacgag gacgatggcg gcagcgagct cgcctcctcc
     2221 gaggacaaca acatggccac catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     2281 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2341 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     2401 ctgtcccccc agttcatgta cggctccgag gcgtacgtga ggcaccccgc cgacatcccc
     2461 gattacaaga agctgccctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgaa
     2521 gacggcggtc tggtgaccgt tacccaggac tcctccctgc aggacggcac gctgatctgc
     2581 aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     2641 atgggctggg aggcctccac cgagatgctg taccccgaag acggcgtgct gaagggccat
     2701 agctatcagg ccctgaagct gaaggacggc ggccactacc tggtggagtt cgagaccatc
     2761 tacatggcca agaagcccgt gcaactgccc ggcgattact gtgtggacac caagctggac
     2821 atcacctccc acaacgagga ctacaccatc gtggaacagt acgggcgctc cgagggccgc
     2881 caccgtctgg gcatggacga gcggtacaag taagaattct gcagatatcc atcacactgg
     2941 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     3001 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3061 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3121 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3181 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3241 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3481 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3601 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3721 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3781 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3901 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3961 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4021 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4081 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4141 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4201 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4261 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4321 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4381 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4441 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4501 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4561 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4621 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4681 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4741 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4801 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4861 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4921 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4981 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5041 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5101 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5161 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5221 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5281 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5341 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5401 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5461 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5521 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5581 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7381 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
