LOCUS       sequence-270943-        5755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S149A 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-S149A
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             698..715
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2635..2756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2672..2691)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2726..2745
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2763..3218)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2900..2921)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3112..3131
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3245..3349
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3351..3708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3351..3542
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3374..3394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3559..3694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3743..4537
                     /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             3743..4537
                     /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(3797..3816)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4407..4426
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4725..4744)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4769..4816
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5145..5733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5634..5653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 ggtttagtga accgtcagat ccgctagcgc taccggtcat catcaccatc accatgctag
      721 cgcttctaac tttactcagt tcgttctcgt cgacaatggc ggaactggcg acgtgactgt
      781 cgccccaagc aacttcgcta acgggatcgc tgaatggatc agctctaact cgcgttcaca
      841 ggcttacaaa gtaacctgta gcgttcgtca gagctctgcg cagaatcgca aatacaccat
      901 caaagtcgag gtacctaaag gcgcctggcg ttcgtactta aatatggaac taaccattcc
      961 aattttcgcc acgaattccg actgcgagct tattgttaag gcaatgcaag gtctcctaaa
     1021 agatggaaac ccgattccct cagcaatcgc agcaaactcc ggcagctaca gatctatggt
     1081 gatggagaag cctagtcccc tgctggtcgg gcgggaattt gtgagacagt attacacact
     1141 gctgaaccag gccccagaca tgctgcatag attttatgga aagaactctt cttatgtcca
     1201 tgggggattg gattcaaatg gaaagccagc agatgcagtc tacggacaga aagaaatcca
     1261 caggaaagtg atgtcacaaa acttcaccaa ctgccacacc aagattcgcc atgttgatgc
     1321 tcatgccacg ctaaatgatg gtgtggtagt ccaggtgatg gggcttctct ctaacaacaa
     1381 ccaggctttg aggagattca tgcaaacgtt tgtccttgct cctgaggggt ctgttgcaaa
     1441 taaattctat gttcacaatg atatcttcag ataccaagat gaggtctttg gtgggtttgt
     1501 cactgagcct caggaggagg ctgaagaaga agtagaggaa cctgaagaaa gacagcaaac
     1561 acctgaggtg gtacctgatg attctggaac tttctatgat caggcagttg tcagtaatga
     1621 catggaagaa catttagagg agcctgttgc tgaaccagag cctgatcctg aaccagaacc
     1681 agaacaagaa cctgtatctg aaatccaaga ggaaaagcct gagccagtat tagaagaaac
     1741 tgcccctgag gatgctcaga agagttcttc tccagcacct gcagacatag ctcagacagt
     1801 acaggaagac ttgaggacat tttcttgggc atctgtgacc agtaagaatc ttccacccag
     1861 tggagctgtt ccagttactg ggataccacc tcatgttgtt aaagtaccag cttcacagcc
     1921 ccgtccagag tctaagcctg aatctcagat tccaccacaa agacctcagc gggatcaaag
     1981 agtgcgagaa caacgaataa atattcctcc ccaaagggga cccagaccaa tccgtgaggc
     2041 tggtgagcaa ggtgacattg aaccccgaag aatggtgaga caccctgaca gtcaccaact
     2101 cttcattggc aacctgcctc atgaagtgga caaatcagag cttaaagatt tctttcaaag
     2161 ttatggaaac gtggtggagt tgcgcattaa cagtggtggg aaattaccca attttggttt
     2221 tgttgtgttt gatgattctg agcctgttca gaaagtcctt agcaacagac ccatcatgtt
     2281 cagaggtgag gtccgtctga atgtcgaaga gaagaagact cgagctgcca gggaaggcga
     2341 ccgacgagat aatcgccttc ggggacctgg aggccctcga ggtgggctgg gtggtggaat
     2401 gagaggccct ccccgtggag gcatggtgca gaaaccagga tttggagtgg gaagggggct
     2461 tgcgccacgg cagggaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct
     2521 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2581 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2641 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2701 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     2761 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2821 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2881 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2941 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3001 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3061 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3121 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3181 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3241 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3301 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3361 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3721 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3781 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3841 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3901 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3961 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4021 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4081 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4141 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4201 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4261 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4321 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4381 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4441 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4501 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4561 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4621 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4681 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4741 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4801 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4861 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4921 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4981 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5041 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5101 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5161 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5221 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5281 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5341 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5401 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5461 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5521 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5581 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5641 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5701 tgatgctcgt caggggggcg gagcctatgg aaaaaggcca gcaacgcggc ctttt
//
