LOCUS       sequence-263066-        5757 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 5757)
  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 5757)
  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..5757
                     /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             606..623
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2545..2666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2582..2601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2636..2655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2673..3128)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2810..2831)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3022..3041
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3155..3259
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3261..3618
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3261..3452
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3284..3304)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3469..3604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3653..4447
                     /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             3653..4447
                     /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(3707..3726)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4317..4336
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4635..4654)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4679..4726
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5055..5643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5544..5563
                     /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 ccggtcatca tcaccatcac catgctagcg cttctaactt tactcagttc gttctcgtcg
      661 acaatggcgg aactggcgac gtgactgtcg ccccaagcaa cttcgctaac gggatcgctg
      721 aatggatcag ctctaactcg cgttcacagg cttacaaagt aacctgtagc gttcgtcaga
      781 gctctgcgca gaatcgcaaa tacaccatca aagtcgaggt acctaaaggc gcctggcgtt
      841 cgtacttaaa tatggaacta accattccaa ttttcgccac gaattccgac tgcgagctta
      901 ttgttaaggc aatgcaaggt ctcctaaaag atggaaaccc gattccctca gcaatcgcag
      961 caaactccgg cagctacaga tctatggtga tggagaagcc tagtcccctg ctggtcgggc
     1021 gggaatttgt gagacagtat tacacactgc tgaaccaggc cccagacatg ctgcatagat
     1081 tttatggaaa gaactcttct tatgtccatg ggggattgga ttcaaatgga aagccagcag
     1141 atgcagtcta cggacagaaa gaaatccaca ggaaagtgat gtcacaaaac ttcaccaact
     1201 gccacaccaa gattcgccat gttgatgctc atgccacgct aaatgatggt gtggtagtcc
     1261 aggtgatggg gcttctctct aacaacaacc aggctttgag gagattcatg caaacgtttg
     1321 tccttgctcc tgaggggtct gttgcaaata aattctatgt tcacaatgat atcttcagat
     1381 accaagatga ggtctttggt gggtttgtca ctgagcctca ggaggagtct gaagaagaag
     1441 tagaggaacc tgaagaaaga cagcaaacac ctgaggtggt acctgatgat tctggaactt
     1501 tctatgatca ggcagttgtc agtaatgaca tggaagaaca tttagaggag cctgttgctg
     1561 aaccagagcc tgatcctgaa ccagaaccag aacaagaacc tgtatctgaa atccaagagg
     1621 aaaagcctga gccagtatta gaagaaactg cccctgagga tgctcagaag agttcttctc
     1681 cagcacctgc agacatagct cagacagtac aggaagactt gaggacattt tcttgggcat
     1741 ctgtgaccag taagaatctt ccacccagtg gagctgttcc agttactggg ataccacctc
     1801 atgttgttaa agtaccagct tcacagcccc gtccagagtc taagcctgaa tctcagattc
     1861 caccacaaag acctcagcgg gatcaaagag tgcgagaaca acgaataaat attcctcccc
     1921 aaaggggacc cagaccaatc cgtgaggctg gtgagcaagg tgacattgaa ccccgaagaa
     1981 tggtgagaca ccctgacagt caccaactct tcattggcaa cctgcctcat gaagtggaca
     2041 aatcagagct taaagatttc tttcaaagtt atggaaacgt ggtggagttg cgcattaaca
     2101 gtggtgggaa attacccaat tttggttttg ttgtgtttga tgattctgag cctgttcaga
     2161 aagtccttag caacaggccc atcatgttca gaggtgaggt ccgtctgaat gtcgaagaga
     2221 agaagactcg agctgccagg gaaggcgacc gacgagataa tcgccttcgg ggacctggag
     2281 gccctcgagg tgggctgggt ggtggaatga gaggccctcc ccgtggaggc atggtgcaga
     2341 aaccaggatt tggagtggga agggggcttg cgccacggca gtgagaattc tgcagtcgac
     2401 ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag ccataccaca
     2461 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2521 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2581 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2641 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     2701 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2761 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2821 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2881 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     2941 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3001 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3061 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3121 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3181 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3241 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3301 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3361 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3421 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3481 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3541 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3601 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3661 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3721 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3781 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     3841 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3901 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3961 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4021 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4081 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4141 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4201 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4261 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4321 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4381 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4441 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4501 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4561 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4621 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4681 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     4741 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4801 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4861 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     4921 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4981 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5041 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5101 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5161 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5221 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5281 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5341 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5401 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5461 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5521 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5581 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5641 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
