LOCUS       sequence-98352-e        8092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent FPX sensor for PIP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8092)
  AUTHORS   Ding Y, Li J, Enterina JR, Shen Y, Zhang I, Tewson PH, Mo GC, Zhang 
            J, Quinn AM, Hughes TE, Maysinger D, Alford SC, Zhang Y, Campbell RE
  TITLE     Ratiometric biosensors based on dimerization-dependent fluorescent 
            protein exchange.
  JOURNAL   Nat Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261.
  PUBMED    25622108
REFERENCE   2  (bases 1 to 8092)
  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"; journalName: "Nat 
            Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261."
COMMENT     Sequence Label: pPHT-PIP2
FEATURES             Location/Qualifiers
     source          1..8092
                     /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"
     misc_feature    1601..1657
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2183..2236
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    2927..2983
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(3517..3534)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3523..3747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3793..4221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3880..3899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4090..4111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4230..4250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4235..4564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4415..4550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4477..4496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4613..5638
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4613..5638
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5768..5889
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5805..5824)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5859..5878
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5938..5954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5938..5954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5951..5973)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5962..5978
                     /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(5986..5992,5993..6010,6011..6016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6031..6052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6169..6186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6340..6925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6420..6439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7096..7887,7888..7956))
                     /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     7719..7738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7957..8061)
                     /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 gctggctagc
      901 gtttaccatg gtgagcaaga gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg
      961 cttggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1021 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     1081 ctgggacatc ctgtcccccc agatcatgta cggctccaag gcgtacgtga agcaccccgc
     1141 cgacgtcccc gattacatga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1201 gcacttcgag gacggcggtc tggtgaccgt gactcaggac acctccctgc aggacggcac
     1261 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     1321 gaagaagacc ttgggctggg attatgccac cgagcgcctg taccccgaag acggcgtgct
     1381 gaagggcgag cttctggggc gcctgaagct gaaggacggc ggcctcaacc tggtggagtt
     1441 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact tcgtggacac
     1501 caagctgggc atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc
     1561 cgagggccgc caccacctgg gcatggacga gctgtacaag agatctcgag ctcaagcttc
     1621 gaattctgca gtcgacggta ccgcgggccc gggatccatg gactcgggcc gggacttcct
     1681 gaccctgcac ggcctacagg atgatgagga tctacaggcg ctgctgaagg gcagccagct
     1741 cctgaaggtg aagtccagct catggaggag agagcgcttc tacaagttgc aggaggactg
     1801 caagaccatc tggcaggagt cccgcaaggt catgcggacc ccggagtccc agctgttctc
     1861 catcgaggac attcaggagg tgcgaatggg gcaccgcacg gagggtctgg agaagttcgc
     1921 ccgtgatgtg cccgaggacc gctgcttctc cattgtcttc aaggaccagc gcaatacact
     1981 agacctcatc gccccatcgc cagctgatgc ccagcactgg gtgctggggc tgcacaagat
     2041 catccaccac tcaggctcca tggaccagcg tcagaagcta cagcactgga ttcactcctg
     2101 cttgcgaaaa gctgacaaaa acaaggacaa caagatgagc ttcaaggagc tgcagaactt
     2161 cctgaaggag ctcaacatcc aggagggcag aggaagtctt ctaacatgcg gtgacgtgga
     2221 ggagaatccc gggcctgtga gcaagggcga ggagaccatc aaagagttca tgcgcttcaa
     2281 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2341 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     2401 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccgaggcgt acgtgaggca
     2461 ccccgccgac atccccgatt acaagaagct gcccttcccc gagggcttca agtgggagcg
     2521 cgtgatgaac ttcgaagacg gcggtctggt gaccgttacc caggactcct ccctgcagga
     2581 cggcacgctg atctgcaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     2641 aatgcagaag aagaccatgg gctgggaggc ctccaccgag atgctgtacc ccgaagacgg
     2701 cgtgctgaag ggccatagct atcaggccct gaagctgaag gacggcggcc actacctggt
     2761 ggagttcgag accatctaca tggccaagaa gcccgtgcaa ctgcccggcg attactgtgt
     2821 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacgg
     2881 gcgctccgag ggccgccacc gtctgggcat ggacgagcgg tacaagagat ctcgagctca
     2941 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccatggact cgggccggga
     3001 cttcctgacc ctgcacggcc tacaggatga tgaggatcta caggcgctgc tgaagggcag
     3061 ccagctcctg aaggtgaagt ccagctcatg gaggagagag cgcttctaca agttgcagga
     3121 ggactgcaag accatctggc aggagtcccg caaggtcatg cggaccccgg agtcccagct
     3181 gttctccatc gaggacattc aggaggtgcg aatggggcac cgcacggagg gtctggagaa
     3241 gttcgcccgt gatgtgcccg aggaccgctg cttctccatt gtcttcaagg accagcgcaa
     3301 tacactagac ctcatcgccc catcgccagc tgatgcccag cactgggtgc tggggctgca
     3361 caagatcatc caccactcag gctccatgga ccagcgtcag aagctacagc actggattca
     3421 ctcctgcttg cgaaaagctg acaaaaacaa ggacaacaag atgagcttca aggagctgca
     3481 gaacttcctg aaggagctca acatccagta gcgcgccctc gactgtgcct tctagttgcc
     3541 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3601 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3661 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3721 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3781 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3841 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3901 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3961 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4021 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4081 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4141 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4201 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4501 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4561 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgatgaaaaa
     4621 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4681 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4741 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4801 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     4861 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4921 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     4981 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     5041 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     5101 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     5161 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     5221 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     5281 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5341 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     5401 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5461 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5521 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     5581 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     5641 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5701 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5761 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5821 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5881 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5941 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6001 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6061 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6121 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6181 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6241 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6301 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6361 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6421 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6481 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6541 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6601 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6661 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6721 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6781 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6841 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7021 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7081 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7141 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7201 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7261 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7321 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7381 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7441 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7501 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7561 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7621 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7681 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7741 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7801 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7861 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7921 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7981 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8041 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
