LOCUS       sequence-270949-        5755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT G3BP1 inserted with MS2BP tagged on the N-terminus to use with 
            the Tethering Assay (MS2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5755)
  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 5755)
  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: pCI-neo-His-MS2BP-G3BP1-WT
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             605..622
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2542..2663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2579..2598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2633..2652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2670..3125)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2807..2828)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3019..3038
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3152..3256
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3258..3615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3258..3449
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3281..3301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3466..3601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3650..4444
                     /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             3650..4444
                     /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(3704..3723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4314..4333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4632..4651)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4676..4723
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5052..5640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5541..5560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggtcatcat caccatcacc atgctagcgc ttctaacttt actcagttcg ttctcgtcga
      661 caatggcgga actggcgacg tgactgtcgc cccaagcaac ttcgctaacg ggatcgctga
      721 atggatcagc tctaactcgc gttcacaggc ttacaaagta acctgtagcg ttcgtcagag
      781 ctctgcgcag aatcgcaaat acaccatcaa agtcgaggta cctaaaggcg cctggcgttc
      841 gtacttaaat atggaactaa ccattccaat tttcgccacg aattccgact gcgagcttat
      901 tgttaaggca atgcaaggtc tcctaaaaga tggaaacccg attccctcag caatcgcagc
      961 aaactccggc agctacagat ctatggtgat ggagaagcct agtcccctgc tggtcgggcg
     1021 ggaatttgtg agacagtatt acacactgct gaaccaggcc ccagacatgc tgcatagatt
     1081 ttatggaaag aactcttctt atgtccatgg gggattggat tcaaatggaa agccagcaga
     1141 tgcagtctac ggacagaaag aaatccacag gaaagtgatg tcacaaaact tcaccaactg
     1201 ccacaccaag attcgccatg ttgatgctca tgccacgcta aatgatggtg tggtagtcca
     1261 ggtgatgggg cttctctcta acaacaacca ggctttgagg agattcatgc aaacgtttgt
     1321 ccttgctcct gaggggtctg ttgcaaataa attctatgtt cacaatgata tcttcagata
     1381 ccaagatgag gtctttggtg ggtttgtcac tgagcctcag gaggagtctg aagaagaagt
     1441 agaggaacct gaagaaagac agcaaacacc tgaggtggta cctgatgatt ctggaacttt
     1501 ctatgatcag gcagttgtca gtaatgacat ggaagaacat ttagaggagc ctgttgctga
     1561 accagagcct gatcctgaac cagaaccaga acaagaacct gtatctgaaa tccaagagga
     1621 aaagcctgag ccagtattag aagaaactgc ccctgaggat gctcagaaga gttcttctcc
     1681 agcacctgca gacatagctc agacagtaca ggaagacttg aggacatttt cttgggcatc
     1741 tgtgaccagt aagaatcttc cacccagtgg agctgttcca gttactggga taccacctca
     1801 tgttgttaaa gtaccagctt cacagccccg tccagagtct aagcctgaat ctcagattcc
     1861 accacaaaga cctcagcggg atcaaagagt gcgagaacaa cgaataaata ttcctcccca
     1921 aaggggaccc agaccaatcc gtgaggctgg tgagcaaggt gacattgaac cccgaagaat
     1981 ggtgagacac cctgacagtc accaactctt cattggcaac ctgcctcatg aagtggacaa
     2041 atcagagctt aaagatttct ttcaaagtta tggaaacgtg gtggagttgc gcattaacag
     2101 tggtgggaaa ttacccaatt ttggttttgt tgtgtttgat gattctgagc ctgttcagaa
     2161 agtccttagc aacagaccca tcatgttcag aggtgaggtc cgtctgaatg tcgaagagaa
     2221 gaagactcga gctgccaggg aaggcgaccg acgagataat cgccttcggg gacctggagg
     2281 ccctcgaggt gggctgggtg gtggaatgag aggccctccc cgtggaggca tggtgcagaa
     2341 accaggattt ggagtgggaa gggggcttgc gccacggcag ggaattctgc agtcgacggt
     2401 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     2461 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2521 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2581 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2641 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     2701 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2761 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2821 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2881 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2941 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3001 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3061 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3121 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3181 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3241 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3481 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3541 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3601 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3661 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3721 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3781 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3841 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3901 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3961 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4021 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4081 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4141 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4201 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4261 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4321 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4381 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4441 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4501 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4561 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4621 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4681 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4741 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4801 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4861 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4921 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcatt
//
