LOCUS       sequence-364041-        7388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded single-color sensor for monitoring plasma 
            membrane PI(4,5)P2 dynamics in living cells. Based on 
            dimerization-dependent green fluorescent protein (ddGFP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7388)
  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 7388)
  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-dPlcR
FEATURES             Location/Qualifiers
     source          1..7388
                     /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(2941..2959)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2942..2959)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2979..2996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2985..3209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3255..3683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3342..3361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3552..3573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3692..3712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3697..4026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3877..4012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3939..3958
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4093..4887
                     /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(4147..4166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4757..4776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5061..5182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5098..5117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5152..5171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5231..5247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5231..5247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5244..5266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5255..5271
                     /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(5279..5285,5286..5303,5304..5309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5324..5345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5462..5479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5633..6221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5713..5732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6392..7183,7184..7252))
                     /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     7015..7034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7253..7357)
                     /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 gcccttacga gggcacccag
     1081 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     1141 cccctgatca tgtacggctc caagatgtac gtgaagcacc ccgccgacgt ccccgattac
     1201 atgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgcactt cgaggacggc
     1261 ggtctggtga ccgcgacaca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg
     1321 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccttgggc
     1381 tgggattatg ccaccgagcg cctgtacccc gaagaaggcg tgctgaaggg cgagctcctg
     1441 gggcgcctga agctgaagga cggcggcctc aacctggtgg agtccaagac catctacatg
     1501 gccaagaagc ccgtgcaact gcccggctac tacttcgtgg acaccaagct ggacatcacc
     1561 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     1621 ctggggcatg gcaccggcag cgcatgcatg gactcgggta gggacttcct gaccctgcac
     1681 gggctccagg atgacccgga ccttcaggcc cttctgaagg gcagccagct tctgaaggtg
     1741 aagtccagct cgtggcgtag ggaacgcttc tacaagctac aggaggactg caagaccatc
     1801 tggcaggaat ctcgaaaggt catgaggtcc ccggagtcgc agctgttctc catcgaggac
     1861 attcaggagg tacggatggg acaccgcaca gaaggcctgg agaagtttgc ccgagacatc
     1921 cccgaggatc gatgcttctc cattgtcttc aaggaccagc gcaacaccct agacctcatt
     1981 gccccatcac cagctgacgc tcagcactgg gtgcagggcc tgcgcaagat catccaccac
     2041 tccggctcca tggaccagcg gcagaagctg cagcactgga ttcactcctg cttgcgaaag
     2101 gctgatgtcg acggcggtag cgtggcagaa gaggaagatg acgaggaaga ggacgaggac
     2161 gatggcggca gcgagctcgc ctcctccgag gacaacaaca tggccaccat caaagagttc
     2221 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     2281 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     2341 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccgaggcg
     2401 tacgtgaggc accccgccga catccccgat tacaagaagc tgcccttccc cgagggcttc
     2461 aagtgggagc gcgtgatgaa cttcgaagac ggcggtctgg tgaccgttac ccaggactcc
     2521 tccctgcagg acggcacgct gatctgcaag gtgaagatgc gcggcaccaa cttccccccc
     2581 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gatgctgtac
     2641 cccgaagacg gcgtgctgaa gggccatagc tatcaggccc tgaagctgaa ggacggcggc
     2701 cactacctgg tggagttcga gaccatctac atggccaaga agcccgtgca actgcccggc
     2761 gattactgtg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     2821 gaacagtacg ggcgctccga gggccgccac cgtctgggca tggacgagcg gtacaagtaa
     2881 gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat
     2941 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3001 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3061 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3121 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3181 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3241 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3301 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3361 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3421 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3481 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3541 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3601 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3661 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3721 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3781 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3841 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3901 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3961 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4021 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4081 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4141 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4201 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4261 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4321 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4381 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4441 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4501 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4561 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4621 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4681 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4741 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4801 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4861 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4921 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4981 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5041 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5101 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5161 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5221 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5281 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5341 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5401 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5461 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5521 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5581 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5641 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5701 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5761 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5821 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5881 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5941 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6001 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6061 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6121 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6481 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6541 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6601 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6661 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6721 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6781 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6841 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6901 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6961 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7021 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7081 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7141 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7201 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7261 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7321 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7381 ctgacgtc
//
