LOCUS       sequence-403346-        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the transmembrane domain from mouse LAT with mEos3.2 fused 
            on the N terminus. The palmitoylation site at C27 is mutated to A. 
            Used as a fluorescent plasma membrane probe. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4888)
  AUTHORS   Shelby SA, Castello-Serrano I, Wisser KC, Levental I, Veatch SL
  TITLE     Membrane phase separation drives responsive assembly of receptor 
            signaling domains.
  JOURNAL   Nat Chem Biol. 2023 Jun;19(6):750-758. doi: 
            10.1038/s41589-023-01268-8. Epub 2023 Mar 30.
  PUBMED    36997644
REFERENCE   2  (bases 1 to 4888)
  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"; doi: 
            "10.1038/s41589-023-01268-8"; journalName: "Nat Chem Biol"; date: 
            "2023-06"; volume: "19"; issue: "6"; pages: "750-758"
COMMENT     Sequence Label: mEos3.2trLAT-P
FEATURES             Location/Qualifiers
     source          1..4888
                     /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      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             718..1395
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    1676..1797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1713..1732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1767..1786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1804..2259)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1941..1962)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2153..2172
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2286..2390
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2392..2749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2392..2583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2415..2435)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2600..2735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2784..3578
                     /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             2784..3578
                     /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(2838..2857)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3448..3467
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3766..3785)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3810..3857
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4186..4774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4675..4694
                     /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 gctagccacc
      601 atggagctct tttggagtat agtctttact gtcctcctga gtttctcctg ccgggggtca
      661 gactgggaat ctctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg
      721 agtgcgatta agccagacat gaagatcaaa ctccgtatgg aaggcaacgt aaacgggcac
      781 cactttgtga tcgacggaga tggtacaggc aagccttttg agggaaaaca gagtatggat
      841 cttgaagtca aagagggcgg acctctgcct tttgcctttg atatcctgac cactgcattc
      901 cattacggca acagggtatt cgccaaatat ccagacaaca tacaagacta ttttaagcag
      961 tcgtttccta aggggtattc gtgggaacga agcttgactt tcgaagacgg gggcatttgc
     1021 aacgccagaa acgacataac aatggaaggg gacactttct ataataaagt tcgattttat
     1081 ggtaccaact ttcccgccaa tggtccagtt atgcagaaga agacgctgaa atgggagccc
     1141 tccactgaga aaatgtatgt gcgtgatgga gtgctgacgg gtgatattga gatggctttg
     1201 ttgcttgaag gaaatgccca ttaccgatgt gacttcagaa ctacttacaa agctaaggag
     1261 aagggtgtca agttaccagg cgcccacttt gtggaccact gcattgagat tttaagccat
     1321 gacaaagatt acaacaaggt taagctgtat gagcatgctg ttgctcattc tggattgcct
     1381 gacaatgcca gacgattctc gagaatggaa gcagacgcct tgagcccggt ggggctgggc
     1441 ctcctgctgc tgcccttctt ggtcacgctc ctggctgccc tggccgtgcg ctgccgtgag
     1501 ttgccagtct cctaagaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     1561 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1621 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1681 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1741 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     1801 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1861 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1921 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1981 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2041 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2101 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2161 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2221 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2401 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2461 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2521 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2581 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2641 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2701 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     2761 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2821 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2881 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2941 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3001 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3061 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3121 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3181 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3241 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3301 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3361 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3421 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3481 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3541 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3601 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3661 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3721 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     3781 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3841 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3901 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3961 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4021 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4081 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4141 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4201 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4261 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4321 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4381 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4441 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4501 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4561 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4621 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4681 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4741 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4801 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4861 tgtggataac cgtattaccg ccatgcat
//
