LOCUS       sequence-270958-        6116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S149A G3BP1 inserted with GFP tagged on the N-terminus used to 
            stably integrate into cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  AUTHORS   Fischer JW, Busa VF, Shao Y, Leung AKL
  TITLE     Structure-Mediated RNA Decay by UPF1 and G3BP1.
  JOURNAL   Mol Cell. 2020 Apr 2;78(1):70-84.e6. doi: 
            10.1016/j.molcel.2020.01.021. Epub 2020 Feb 3.
  PUBMED    32017897
REFERENCE   2  (bases 1 to 6116)
  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: 
            "10.1016/j.molcel.2020.01.021"; journalName: "Mol Cell"; date: 
            "2020-04-2- 2"; volume: "78"; issue: "1"; pages: "70-84.e6"
COMMENT     Sequence Label: pEGFP-C1-G3BP1-S149A
FEATURES             Location/Qualifiers
     source          1..6116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        124..427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        428..631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     582..602
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(676..678,679..681,682..1392)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(676..678,679..681,682..1392)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(721..742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(982..1001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1329..1350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2967..3088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3004..3023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3058..3077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3095..3550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3232..3253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3444..3463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3577..3681
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3683..4040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3683..3874
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3706..3726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3891..4026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4075..4869
                     /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             4075..4869
                     /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(4129..4148)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4739..4758
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5057..5076)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5101..5148
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5477..6065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5966..5985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
       61 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      601 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      661 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      781 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      841 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1081 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1141 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1201 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1261 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1321 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1381 gagctgtaca agtccggact cagatctatg gtgatggaga agcctagtcc cctgctggtc
     1441 gggcgggaat ttgtgagaca gtattacaca ctgctgaacc aggccccaga catgctgcat
     1501 agattttatg gaaagaactc ttcttatgtc catgggggat tggattcaaa tggaaagcca
     1561 gcagatgcag tctacggaca gaaagaaatc cacaggaaag tgatgtcaca aaacttcacc
     1621 aactgccaca ccaagattcg ccatgttgat gctcatgcca cgctaaatga tggtgtggta
     1681 gtccaggtga tggggcttct ctctaacaac aaccaggctt tgaggagatt catgcaaacg
     1741 tttgtccttg ctcctgaggg gtctgttgca aataaattct atgttcacaa tgatatcttc
     1801 agataccaag atgaggtctt tggtgggttt gtcactgagc ctcaggagga ggctgaagaa
     1861 gaagtagagg aacctgaaga aagacagcaa acacctgagg tggtacctga tgattctgga
     1921 actttctatg atcaggcagt tgtcagtaat gacatggaag aacatttaga ggagcctgtt
     1981 gctgaaccag agcctgatcc tgaaccagaa ccagaacaag aacctgtatc tgaaatccaa
     2041 gaggaaaagc ctgagccagt attagaagaa actgcccctg aggatgctca gaagagttct
     2101 tctccagcac ctgcagacat agctcagaca gtacaggaag acttgaggac attttcttgg
     2161 gcatctgtga ccagtaagaa tcttccaccc agtggagctg ttccagttac tgggatacca
     2221 cctcatgttg ttaaagtacc agcttcacag ccccgtccag agtctaagcc tgaatctcag
     2281 attccaccac aaagacctca gcgggatcaa agagtgcgag aacaacgaat aaatattcct
     2341 ccccaaaggg gacccagacc aatccgtgag gctggtgagc aaggtgacat tgaaccccga
     2401 agaatggtga gacaccctga cagtcaccaa ctcttcattg gcaacctgcc tcatgaagtg
     2461 gacaaatcag agcttaaaga tttctttcaa agttatggaa acgtggtgga gttgcgcatt
     2521 aacagtggtg ggaaattacc caattttggt tttgttgtgt ttgatgattc tgagcctgtt
     2581 cagaaagtcc ttagcaacag acccatcatg ttcagaggtg aggtccgtct gaatgtcgaa
     2641 gagaagaaga ctcgagctgc cagggaaggc gaccgacgag ataatcgcct tcggggacct
     2701 ggaggccctc gaggtgggct gggtggtgga atgagaggcc ctccccgtgg aggcatggtg
     2761 cagaaaccag gatttggagt gggaaggggg cttgcgccac ggcagggaat tctgcagtcg
     2821 acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     2881 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2941 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3061 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3121 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3181 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3241 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3301 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3361 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3421 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3481 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3541 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3661 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3961 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4021 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5041 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5101 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5161 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5221 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5281 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5341 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5401 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6061 ggaaaaaggc cagcaacgcg gcctttttac ggttcctggc cttttgctgg cctttt
//
