LOCUS       sequence-269246-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G3BP1 shRNA (Targeting 3'UTR) inserted into the PLKO.1 plasmid 
            (GCCTGTAAGAAATACAGGATT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  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 7084)
  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: PLKO.1-G3BP1-3'UTR
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        116..364
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     116..136
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     287..306
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    461..578
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        627..1137
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1056..1074
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1159..1758
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1159..1178)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1655..1675
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1886..2119
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2191..2312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2228..2247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2282..2301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2352..2487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2507..2526)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2508..2526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2536..2553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2536..2552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2545..2567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2694..3149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2781..2800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2991..3012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3175..3279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3280..3348,3349..4140)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3498..3517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4311..4899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4800..4819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5053..5070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5187..5208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5223..5228,5229..5246,5247..5253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5261..5277
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5266..5288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5285..5301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5285..5301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5320..5340
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5322..5340
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5368..5594
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5595..5775
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5822..5947
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6440..6673
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6858..6902
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
       61 cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg
      121 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
      181 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
      241 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
      301 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
      361 caccgggcct gtaagaaata caggattctc gagaatcctg tatttcttac aggctttttg
      421 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
      481 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      541 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      601 gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct
      661 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
      721 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
      781 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
      841 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
      901 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
      961 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
     1021 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
     1081 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
     1141 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     1201 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     1261 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     1321 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     1381 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     1441 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     1501 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     1561 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     1621 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     1681 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     1741 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     1801 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     1861 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     1921 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     1981 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2041 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2101 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     2161 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     2221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     2341 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2401 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2461 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     2521 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2581 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2641 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     2701 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2761 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2821 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2881 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2941 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3001 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3061 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3121 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3961 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4441 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5101 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5161 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     5221 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5281 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     5341 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     5401 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     5461 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     5521 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     5581 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5641 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5701 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5761 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     5821 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     5881 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     5941 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     6001 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     6061 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     6121 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     6181 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     6241 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     6301 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     6361 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     6421 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     6481 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     6541 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     6601 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     6661 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     6721 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     6781 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     6841 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     6901 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     6961 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     7021 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     7081 ggcc
//
