LOCUS       sequence-403342-        4856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the transmembrane domain from mouse LAT with mEos3.2 fused 
            on the N terminus. Palmitoylation sites at C27 and C30 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 4856)
  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 4856)
  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-2P
FEATURES             Location/Qualifiers
     source          1..4856
                     /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    1644..1765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1681..1700)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1735..1754
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1772..2227)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1909..1930)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2121..2140
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2254..2358
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2360..2717
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2360..2551
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2383..2403)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2568..2703
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2752..3546
                     /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             2752..3546
                     /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(2806..2825)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3416..3435
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3734..3753)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3778..3825
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4154..4742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4643..4662
                     /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 cgcccgtgag
     1501 ttgccagtct cctaagcggc cgcgactcta gataactgat cataatcagc cataccacat
     1561 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1621 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1681 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1741 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     1801 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1861 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     1921 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1981 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2041 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2101 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2161 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2221 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2341 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2401 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2461 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2521 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2581 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2641 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2701 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     2761 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2821 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2881 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     2941 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3001 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3061 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3121 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3181 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3241 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3301 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3361 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3421 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3481 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3541 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3601 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3661 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     3721 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3781 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     3841 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     3901 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     3961 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4021 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4081 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4261 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4801 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
