LOCUS       sequence-399534-        6219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses fluorescent protein-tagged lipid biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  AUTHORS   Wills RC, Doyle CP, Zewe JP, Pacheco J, Hansen SD, Hammond GRV
  TITLE     A novel homeostatic mechanism tunes PI(4,5)P2-dependent signaling at
            the plasma membrane.
  JOURNAL   J Cell Sci. 2023 Aug 3:jcs.261494. doi: 10.1242/jcs.261494.
  PUBMED    37534432
REFERENCE   2  (bases 1 to 6219)
  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: "J Cell Sci.
            2023 Aug 3:jcs.261494. doi: 10.1242/jcs.261494."
COMMENT     Sequence Label: PH-PLCd1-mNGx3
FEATURES             Location/Qualifiers
     source          1..6219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..134
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     85..105
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             728..1435
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      983..992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1445..1454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1451..2158
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1706..1715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2168..2177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2174..2884
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2429..2438
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3007..3128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3098..3117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3135..3590)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3272..3293)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3484..3503
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3617..3721
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3723..4080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3723..3914
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3746..3766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3931..4066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4115..4909
                     /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             4115..4909
                     /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(4169..4188)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4779..4798
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5097..5116)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5141..5188
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5517..6105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6006..6025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
       61 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      121 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      181 cgagctcaag cttcgaattc acggcatgga ctcgggccgg gacttcctga ccctgcacgg
      241 cctacaggat gatgaggatc tacaggcgct gctgaagggc agccagctcc tgaaggtgaa
      301 gtccagctca tggaggagag agcgcttcta caagttgcag gaggactgca agaccatctg
      361 gcaggagtcc cgcaaggtca tgcggacccc ggagtcccag ctgttctcca tcgaggacat
      421 tcaggaggtg cgaatggggc accgcacgga gggtctggag aagttcgccc gtgatgtgcc
      481 cgaggaccgc tgcttctcca ttgtcttcaa ggaccagcgc aatacactag acctcatcgc
      541 cccatcgcca gctgatgccc agcactgggt gctggggctg cacaagatca tccaccactc
      601 aggctccatg gaccagcgtc agaagctaca gcactggatt cactcctgct tgcgaaaagc
      661 tgacaaaaac aaggacaaca agatgagctt caaggagctg cagaacttcc tgaagggggt
      721 gggtggcatg gtgagcaagg gcgaggagga taacatggcc tctctcccag cgacacatga
      781 gttacacatc tttggctcca tcaacggtgt ggactttgac atggtgggtc agggcaccgg
      841 caatccaaat gatggttatg aggagttaaa cctgaagtcc accaagggtg acctccagtt
      901 ctccccctgg attctggtcc ctcatatcgg gtatggcttc catcagtacc tgccctaccc
      961 tgacgggatg tcgcctttcc aggccgccat ggtagatggc tccggatacc aagtccatcg
     1021 cacaatgcag tttgaagatg gtgcctccct tactgttaac taccgctaca cctacgaggg
     1081 aagccacatc aaaggagagg cccaggtgaa ggggactggt ttccctgctg acggtcctgt
     1141 gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg aagaagactt accccaacga
     1201 caaaaccatc atcagtacct ttaagtggag ttacaccact ggaaatggca agcgctaccg
     1261 gagcactgcg cggaccacct acacctttgc caagccaatg gcggctaact atctgaagaa
     1321 ccagccgatg tacgtgttcc gtaagacgga gctcaagcac tccaagaccg agctcaactt
     1381 caaggagtgg caaaaggcct ttaccgatgt gatgggcatg gacgagctgt acaagtcacc
     1441 ggtcgccacc atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca
     1501 tgagttacac atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac
     1561 cggcaatcca aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca
     1621 gttctccccc tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta
     1681 ccctgacggg atgtcgcctt tccaggccgc catggtagat ggctccggat accaagtcca
     1741 tcgcacaatg cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga
     1801 gggaagccac atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc
     1861 tgtgatgacc aactcgctga ccgctgcgga ctggtgcagg tcgaagaaga cttaccccaa
     1921 cgacaaaacc atcatcagta cctttaagtg gagttacacc actggaaatg gcaagcgcta
     1981 ccggagcact gcgcggacca cctacacctt tgccaagcca atggcggcta actatctgaa
     2041 gaaccagccg atgtacgtgt tccgtaagac ggagctcaag cactccaaga ccgagctcaa
     2101 cttcaaggag tggcaaaagg cctttaccga tgtgatgggc atggacgagc tgtacaagtc
     2161 accggtcgcc accatggtga gcaagggcga ggaggataac atggcctctc tcccagcgac
     2221 acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg
     2281 caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct
     2341 ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc
     2401 ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt
     2461 ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta
     2521 cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg
     2581 tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc
     2641 caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg
     2701 ctaccggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct
     2761 gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct
     2821 caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa
     2881 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     2941 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3001 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3061 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3121 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3181 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3241 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3301 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3361 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3421 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3481 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3541 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3721 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4081 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4141 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4201 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4261 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4321 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4381 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4441 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4501 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4561 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4621 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4681 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4741 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4801 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4861 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4921 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4981 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5041 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5101 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5161 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5221 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5281 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5341 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5401 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5461 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5521 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5581 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5641 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5701 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5761 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5821 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5881 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5941 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6001 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6061 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6121 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6181 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
