LOCUS       sequence-403337-        4698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the membrane anchor of NRas, including the last 10 residues 
            of the C terminus with 1 farnesylation and 1 palmitoylation, with 
            mEos3.2 fused to the N terminus. Used as a fluorescent plasma 
            membrane probe. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4698)
  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 4698)
  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: mEos3.2FP
FEATURES             Location/Qualifiers
     source          1..4698
                     /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"
     CDS             613..1290
                     /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    1486..1607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1523..1542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1577..1596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1614..2069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1751..1772)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1963..1982
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2096..2200
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2202..2559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2202..2393
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2225..2245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2410..2545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2594..3388
                     /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             2594..3388
                     /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(2648..2667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3258..3277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3576..3595)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3620..3667
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3996..4584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4485..4504
                     /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 ccggtcgcca ccatgagtgc gattaagcca gacatgaaga tcaaactccg tatggaaggc
      661 aacgtaaacg ggcaccactt tgtgatcgac ggagatggta caggcaagcc ttttgaggga
      721 aaacagagta tggatcttga agtcaaagag ggcggacctc tgccttttgc ctttgatatc
      781 ctgaccactg cattccatta cggcaacagg gtattcgcca aatatccaga caacatacaa
      841 gactatttta agcagtcgtt tcctaagggg tattcgtggg aacgaagctt gactttcgaa
      901 gacgggggca tttgcaacgc cagaaacgac ataacaatgg aaggggacac tttctataat
      961 aaagttcgat tttatggtac caactttccc gccaatggtc cagttatgca gaagaagacg
     1021 ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg atggagtgct gacgggtgat
     1081 attgagatgg ctttgttgct tgaaggaaat gcccattacc gatgtgactt cagaactact
     1141 tacaaagcta aggagaaggg tgtcaagtta ccaggcgccc actttgtgga ccactgcatt
     1201 gagattttaa gccatgacaa agattacaac aaggttaagc tgtatgagca tgctgttgct
     1261 cattctggat tgcctgacaa tgccagacga ttctcgagct caagcttgaa ttctgcagtc
     1321 gacggttgta tgggattgcc atgtgtggtg atgtaaggat ccaccggatc tagataactg
     1381 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1441 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1501 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1561 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     1621 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1681 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1741 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1801 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1861 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1921 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1981 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2041 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2161 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2221 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2341 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2401 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2461 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2521 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2581 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2641 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2701 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2761 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2821 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2881 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2941 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3001 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3061 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3121 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3181 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3241 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3301 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3361 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3421 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3481 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3541 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3601 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3661 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3721 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3781 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3841 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3901 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3961 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4021 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4081 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4141 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4201 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4261 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4321 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4381 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4441 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4501 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4561 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4621 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4681 cgtattaccg ccatgcat
//
