LOCUS       sequence-399277-        5193 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 5193)
  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 5193)
  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-P4C
FEATURES             Location/Qualifiers
     source          1..5193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        349..552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     503..523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      585..594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             591..1298
                     /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     complement(736..754)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1965..2086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2002..2021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2056..2075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2093..2548)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2230..2251)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2442..2461
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2575..2679
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2681..3038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2681..2872
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2704..2724)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2889..3024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3073..3867
                     /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             3073..3867
                     /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(3127..3146)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3737..3756
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4055..4074)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4099..4146
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4475..5063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4964..4983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
       61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      121 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      181 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      241 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      301 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      541 ataagcagag ctggtttagt gaaccgtcag atccgctagc ggtcgccacc atggtgagca
      601 agggcgagga ggataacatg gccatcatta aggagttcat gcgcttcaag gtgcacatgg
      661 aggggtccgt gaacggccac gtgttcgaga tcgagggcga gggcgagggc cgcccctacg
      721 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcacctggg
      781 acatcctgag ccctcagttc atgtacggct ccaatgccta cgtgaagcac cccgccgaca
      841 tccccgacta ctttaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaaat
      901 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
      961 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgtg atgcagaaga
     1021 agaccatggg ctgggaggcc ctctccgagc ggatgtaccc cgaggacggc gccctgaagg
     1081 gcgaggtcaa gcccagagtg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     1141 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aaccgcaagc
     1201 tggacatcac cagccacaac gaggactaca ccatcgtgga gcagtacgag agagccgagg
     1261 gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc
     1321 aagcttcgaa ttcaatgtca aaatattcct ccaagccatt attggatgtg gaattaaata
     1381 aaatcgctga aggattagaa ctcactgcaa aaatttacaa tgaaaagaga ggacgcgaat
     1441 ggtggtttaa aggttcaaga aatgaagctc gtaagaccca atgtgaagaa ttgcaaagag
     1501 tatccaaaga aatcaatact cttctgcaaa gtgaatcttt aacgaaaagc caggtacttg
     1561 aaaaggtttt aaattctata gaaacattag ataaaattga cagagacatt tctgccgaat
     1621 ccaattggtt tcaaagtact ctgcaaaagg aagtcaggtt atttcgagat caattgaaag
     1681 atatttgcca attggacaag tatgccttta aatcaacaaa acttgatgaa atcatctctc
     1741 tggaaatgga agaacaattt caaaagatac aagatcctgc tgttcaacaa attgtcaggg
     1801 acttgccttc tcattgccac aatgatgaag caattggatc caccggatct agataactga
     1861 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1921 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     1981 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2041 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     2101 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2161 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2221 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2281 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2341 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2401 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2461 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2521 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2581 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2641 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     2701 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2761 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2881 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2941 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3001 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3061 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3121 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3181 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3241 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3301 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3361 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3421 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3481 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3541 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3601 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3661 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3721 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3781 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3841 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3901 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3961 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4021 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4081 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4141 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4201 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     4261 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     4321 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4381 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4441 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4501 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4561 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4621 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4681 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4741 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4801 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4861 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4921 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4981 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5041 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5101 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5161 gtattaccgc catgcattag ttattaatag taa
//
