LOCUS       sequence-399535-        5493 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 5493)
  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 5493)
  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-N1-Tubbyc
FEATURES             Location/Qualifiers
     source          1..5493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1443..1452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1449..2159
                     /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(1594..1612)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2281..2402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2318..2337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2372..2391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2409..2864)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2546..2567)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2758..2777
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2891..2995
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2997..3354
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2997..3188
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3020..3040)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3205..3340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3389..4183
                     /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             3389..4183
                     /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(3443..3462)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4053..4072
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4371..4390)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4415..4462
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4791..5379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5280..5299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct atggtggata ttgaggtcca ggatctagag gagtttgcac
      661 tgaggccagc cccacaaggg atcaccatca aatgccgcat cactcgggac aagaagggga
      721 tggaccgcgg catgtacccc acctactttc tgcacctaga ccgtgaggat ggcaagaagg
      781 tgttcctcct ggcgggcagg aagagaaaga agagtaaaac ttccaattac ctcatctctg
      841 tggacccaac agacttgtct cggggaggcg atagctatat cgggaaattg cgatccaacc
      901 tgatgggcac caagttcacc gtttatgaca atggcgtcaa ccctcagaag gcatcctctt
      961 ccacgctgga aagcggaacc ttgcgccagg agctggcagc ggtgtgctat gagacaaatg
     1021 tcctaggctt caagggacct cggaagatga gtgtgatcgt cccaggcatg aacatggttc
     1081 atgagagagt ctgtatccgc ccccgcaatg aacatgagac cctgttagca cgctggcaga
     1141 acaagaacac ggagagcatc attgagctgc agaacaagac gccagtctgg aatgatgaca
     1201 cacagtccta tgtacttaac ttccacggcc gtgtcacaca ggcttctgtg aagaacttcc
     1261 agatcatcca cggcaatgac ccggactaca tcgtcatgca gtttggccgg gtagcagaag
     1321 atgtgttcac catggattac aactacccac tgtgtgcact gcaggccttt gccattgctc
     1381 tgtccagctt tgacagcaag ctggcctgcg agacggtacc gcgggcccgg gatccaccgg
     1441 tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcattaag gagttcatgc
     1501 gcttcaaggt gcacatggag gggtccgtga acggccacgt gttcgagatc gagggcgagg
     1561 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     1621 ccctgccctt cacctgggac atcctgagcc ctcagttcat gtacggctcc aatgcctacg
     1681 tgaagcaccc cgccgacatc cccgactact ttaagctgtc cttccccgag ggcttcaagt
     1741 gggagcgcgt gatgaaattc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1801 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     1861 gccccgtgat gcagaagaag accatgggct gggaggccct ctccgagcgg atgtaccccg
     1921 aggacggcgc cctgaagggc gaggtcaagc ccagagtgaa gctgaaggac ggcggccact
     1981 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2041 acaacgtcaa ccgcaagctg gacatcacca gccacaacga ggactacacc atcgtggagc
     2101 agtacgagag agccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     2161 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2221 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2401 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2461 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2521 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2581 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2641 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2701 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2761 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2821 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2941 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3001 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3061 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3121 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3181 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3241 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3301 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3361 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3421 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3481 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3541 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3601 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3661 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3721 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3781 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3841 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3901 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3961 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4021 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4081 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4141 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4201 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4261 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4321 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4381 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4441 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4501 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4561 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4621 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4681 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4741 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4801 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4861 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4921 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4981 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5041 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5101 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5161 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5221 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5281 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5341 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5401 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5461 gattctgtgg ataaccgtat taccgccatg cat
//
