LOCUS       sequence-399218-        5185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoactivatable lipid biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5185)
  AUTHORS   Pacheco J, Cassidy AC, Zewe JP, Wills RC, Hammond GRV
  TITLE     PI(4,5)P2 diffuses freely in the plasma membrane even within 
            high-density effector protein complexes.
  JOURNAL   J Cell Biol. 2023 Feb 6;222(2):e202204099. doi: 
            10.1083/jcb.202204099. Epub 2022 Nov 23.
  PUBMED    36416724
REFERENCE   2  (bases 1 to 5185)
  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 
            Biol."; date: "2023-02-6"; volume: "222(2):e202204099. doi"; pages: 
            " 10.1083/jcb.202204099. Epub 2022 Nov 23"
COMMENT     Sequence Label: PAmCherry1-C1-LACT-C2
FEATURES             Location/Qualifiers
     source          1..5185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     206..226
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(251..355)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      382..837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(469..488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     679..700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    844..965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(855..874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     909..928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1624..2331)
                     /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     2168..2186
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2328..2337
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2370..2573)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2399..2419)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2574..2877
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3052..3640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3132..3151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3969..4016
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4041..4060
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4248..5042)
                     /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             complement(4248..5042)
                     /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(4359..4378)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4969..4988
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5077..5185,1..249))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5091..5185,1..41)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
       61 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      121 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
      181 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
      241 ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      301 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      361 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      421 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      481 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      541 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      601 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      661 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      721 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      781 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg
      841 cgttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      901 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      961 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1021 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat cagttatcta
     1081 gatccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcctaac agcccagcag
     1141 ctccactcgc agggtgatac ggttgtgcca ggccacgggc tggatccgca cgaagcgagc
     1201 ctggaacggc gtctcaaata tgttcttctt gtgggaatta ttgtccatgt taccagggaa
     1261 aatcttgctt tctgaggccc ccgggtcctt gtactcagtc caggtcacac catcatcacc
     1321 ataggccacc ctgtaggcag ccacatattg aatgtggcca aagtctcggg caccctgggt
     1381 gatgatgccc gtgactcgct tctgggagcc caggtcaatc tgcagccact cagaggcact
     1441 gttggtctgg gcggtccagg cgttgaactt gccctgatta tccagtcgtg cgtagtaggg
     1501 aaaccagcta aaggcactca ggccccaggt tttgtagtag ctggaggctg tgatctgctt
     1561 gttggggatg gtattatcct tcaggcctag gggttcagtg caagatctag atctgagtcc
     1621 ggacttgtac agctcgtcca tgccgccggt ggagtggcgg ccctcggctc tctcgtactg
     1681 ctccacgatg gtgtagtcct cgttgtggct ggtgatgtcc agcttgcggt tgacgttgta
     1741 ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta
     1801 gtggccgccg tccttcagct tcactctggg cttgacctcg cccttcaggg cgccgtcctc
     1861 ggggtacatc cgctcggaga gggcctccca gcccatggtc ttcttctgca tcacggggcc
     1921 gtcggagggg aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag
     1981 ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg aatttcatca cgcgctccca
     2041 cttgaagccc tcggggaagg acagcttaaa gtagtcgggg atgtcggcgg ggtgcttcac
     2101 gtaggcattg gagccgtaca tgaactgagg gctcaggatg tcccaggtga agggcagggg
     2161 gccgcccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc
     2221 ctcgccctcg atctcgaaca cgtggccgtt cacggacccc tccatgtgca ccttgaagcg
     2281 catgaactcc ttaatgatgg ccatgttatc ctcctcgccc ttgctcacca tggtggcgac
     2341 cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca
     2401 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     2461 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     2521 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     2581 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     2641 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     2701 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     2761 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     2821 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     2881 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactaatg
     2941 catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3001 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3061 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3121 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3181 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3241 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3301 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3361 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3421 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3481 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3541 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3601 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3661 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3721 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3781 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     3841 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     3901 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     3961 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     4021 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     4081 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     4141 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     4201 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     4261 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     4321 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     4381 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     4441 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     4501 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     4561 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     4621 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     4681 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     4741 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     4801 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     4861 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     4921 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     4981 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     5041 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     5101 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     5161 aataaaaaaa attagtcagc catgg
//
