LOCUS       sequence-269147-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G3BP1 shRNA (Targeting CDS) inserted into the PLKO.1 plasmid 
            (CGGGAATTTGTGAGACAGTAT).
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-CDS
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..54
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    171..192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(207..212,213..230,231..237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    245..261
                     /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     250..272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     269..285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     269..285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     304..324
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        306..324
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        352..578
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             579..759
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    806..931
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1424..1657
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1842..1886
                     /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"
     promoter        2184..2432
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2184..2204
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2355..2374
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2529..2646
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2695..3205
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3124..3142
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3227..3826
                     /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(3227..3246)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3723..3743
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3954..4187
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4259..4380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4296..4315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4350..4369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4420..4555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4575..4594)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4576..4594)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4604..4621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4604..4620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4613..4635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4762..5217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4849..4868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5059..5080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5243..5347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5348..5416,5417..6208)
                     /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(5566..5585)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6379..6967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6868..6887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
       61 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
      121 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      181 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      241 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
      301 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
      361 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
      421 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
      481 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
      541 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
      601 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      661 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      721 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      781 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      841 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      901 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      961 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     1021 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1081 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1141 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1201 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1261 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     1321 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     1381 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     1441 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     1501 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     1561 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     1621 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     1681 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     1741 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     1801 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     1861 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     1921 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     1981 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     2041 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     2101 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     2161 ctcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
     2221 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     2281 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     2341 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     2401 ctttatatat cttgtggaaa ggacgaaaca ccggcgggaa tttgtgagac agtatctcga
     2461 gatactgtct cacaaattcc cgtttttgaa ttctcgacct cgagacaaat ggcagtattc
     2521 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2581 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2641 aattttcggg tttattacag ggacagcaga gatccacttt ggccgcggct cgagggggtt
     2701 ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg
     2761 tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg
     2821 cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc
     2881 gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc
     2941 cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg
     3001 accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg
     3061 ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc
     3121 gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttga
     3181 ccgaatcacc gacctctctc cccaggggga tccaccggag cttaccatga ccgagtacaa
     3241 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3301 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3361 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3421 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3481 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3541 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3601 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3661 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3721 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3781 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     3841 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcggtacctt taagaccaat
     3901 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     3961 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     4021 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     4081 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     4141 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     4201 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     4261 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4321 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4381 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     4441 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4501 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     4561 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     4621 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4681 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4741 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5161 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5221 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5281 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5341 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5401 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5461 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5521 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5581 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5641 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5701 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5761 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5821 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5881 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5941 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6001 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6061 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6121 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6181 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6241 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6301 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7021 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7081 gtga
//
