LOCUS       sequence-489238-        6116 bp    DNA     circular SYN 14-OCT-2025
DEFINITION  WT 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 Oct 14, 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-WT
FEATURES             Location/Qualifiers
     source          1..6116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..248
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     205..225
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(250..354)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      381..836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(468..487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     678..699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    843..964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(854..873)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     908..927
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2539..3249,3250..3252,3253..3255))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2539..3249,3250..3252,3253..3255))
                     /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(2581..2602)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2930..2949
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3189..3210
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3252..3261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3300..3503)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3329..3349)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3504..3807
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3982..4570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4062..4081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4899..4946
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4971..4990
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5178..5972)
                     /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             complement(5178..5972)
                     /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(5289..5308)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5899..5918
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6007..6116,1..248))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6021..6116,1..40)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
       61 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
      121 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      181 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
      241 cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      361 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
      421 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
      481 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
      541 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
      601 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      661 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      721 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      781 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc
      841 gttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      901 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      961 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1021 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag
     1081 atccggtgga tcccgggccc gcggtaccgt cgactgcaga attccctgcc gtggcgcaag
     1141 cccccttccc actccaaatc ctggtttctg caccatgcct ccacggggag ggcctctcat
     1201 tccaccaccc agcccacctc gagggcctcc aggtccccga aggcgattat ctcgtcggtc
     1261 gccttccctg gcagctcgag tcttcttctc ttcgacattc agacggacct cacctctgaa
     1321 catgatgggt ctgttgctaa ggactttctg aacaggctca gaatcatcaa acacaacaaa
     1381 accaaaattg ggtaatttcc caccactgtt aatgcgcaac tccaccacgt ttccataact
     1441 ttgaaagaaa tctttaagct ctgatttgtc cacttcatga ggcaggttgc caatgaagag
     1501 ttggtgactg tcagggtgtc tcaccattct tcggggttca atgtcacctt gctcaccagc
     1561 ctcacggatt ggtctgggtc ccctttgggg aggaatattt attcgttgtt ctcgcactct
     1621 ttgatcccgc tgaggtcttt gtggtggaat ctgagattca ggcttagact ctggacgggg
     1681 ctgtgaagct ggtactttaa caacatgagg tggtatccca gtaactggaa cagctccact
     1741 gggtggaaga ttcttactgg tcacagatgc ccaagaaaat gtcctcaagt cttcctgtac
     1801 tgtctgagct atgtctgcag gtgctggaga agaactcttc tgagcatcct caggggcagt
     1861 ttcttctaat actggctcag gcttttcctc ttggatttca gatacaggtt cttgttctgg
     1921 ttctggttca ggatcaggct ctggttcagc aacaggctcc tctaaatgtt cttccatgtc
     1981 attactgaca actgcctgat catagaaagt tccagaatca tcaggtacca cctcaggtgt
     2041 ttgctgtctt tcttcaggtt cctctacttc ttcttcagac tcctcctgag gctcagtgac
     2101 aaacccacca aagacctcat cttggtatct gaagatatca ttgtgaacat agaatttatt
     2161 tgcaacagac ccctcaggag caaggacaaa cgtttgcatg aatctcctca aagcctggtt
     2221 gttgttagag agaagcccca tcacctggac taccacacca tcatttagcg tggcatgagc
     2281 atcaacatgg cgaatcttgg tgtggcagtt ggtgaagttt tgtgacatca ctttcctgtg
     2341 gatttctttc tgtccgtaga ctgcatctgc tggctttcca tttgaatcca atcccccatg
     2401 gacataagaa gagttctttc cataaaatct atgcagcatg tctggggcct ggttcagcag
     2461 tgtgtaatac tgtctcacaa attcccgccc gaccagcagg ggactaggct tctccatcac
     2521 catagatctg agtccggact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     2581 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga
     2641 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     2701 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     2761 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     2821 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     2881 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     2941 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     3001 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     3061 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     3121 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     3181 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     3241 gcccttgctc accatggtgg cgaccggtag cgctagcgga tctgacggtt cactaaacca
     3301 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     3361 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     3421 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     3481 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     3541 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     3601 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     3661 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     3721 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     3781 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     3841 cccgtaattg attactatta ataactaatg catggcggta atacggttat ccacagaatc
     3901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     4741 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     4801 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     4861 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     4921 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     4981 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     5041 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     5101 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     5161 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     5221 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     5281 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     5341 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     5401 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     5461 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     5521 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     5581 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     5641 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5701 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5761 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     5821 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5881 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5941 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     6001 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     6061 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggg
//
