LOCUS       sequence-244448-        7098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knock-down of human IRF3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7098)
  AUTHORS   Gratia M, Rodero MP, Conrad C, Bou Samra E, Maurin M, Rice GI, Duffy
            D, Revy P, Petit F, Dale RC, Crow YJ, Amor-Gueret M, Manel N
  TITLE     Bloom syndrome protein restrains innate immune sensing of 
            micronuclei by cGAS.
  JOURNAL   J Exp Med. 2019 May 6;216(5):1199-1213. doi: 10.1084/jem.20181329. 
            Epub 2019 Apr 1.
  PUBMED    30936263
REFERENCE   2  (bases 1 to 7098)
  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: 
            "10.1084/jem.20181329"; journalName: "J Exp Med"; date: "2019-05-6- 
            6"; volume: "216"; issue: "5"; pages: "1199-1213"
COMMENT     Sequence Label: pLKO.1-puro-irf3-shrna5
FEATURES             Location/Qualifiers
     source          1..7098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /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        1840..2088
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1840..1860
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2011..2030
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     ncRNA           2115..2129
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    2199..2316
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2365..2875
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2794..2812
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2897..3496
                     /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(2897..2916)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3393..3413
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3624..3857
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3929..4050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3966..3985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4020..4039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4090..4225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4245..4264)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4246..4264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4274..4291)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4274..4290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4283..4305)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4432..4887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4519..4538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4729..4750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4913..5017
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5018..5086,5087..5878)
                     /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(5236..5255)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6049..6637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6538..6557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6791..6808
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6925..6946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6961..6966,6967..6984,6985..6991)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6999..7015
                     /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     7004..7026
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7023..7039
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7023..7039
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7058..7078
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7060..7078
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cgatcacgag actagcctcg agcggccgcc cccttcaccg agggcctatt tcccatgatt
     1861 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
     1921 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     1981 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     2041 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg tacccaggaa
     2101 gacattctgg atctgtgaag ccacagatgg gatccagaat gtcttcctgg gtatttttaa
     2161 ttctcgacct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2221 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2281 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2341 gatccacttt ggccgcggct cgagggggtt ggggttgcgc cttttccaag gcagccctgg
     2401 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
     2461 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
     2521 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
     2581 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
     2641 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
     2701 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
     2761 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
     2821 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggggga
     2881 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     2941 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3001 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3061 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3121 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3181 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3241 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3301 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3361 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3421 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3481 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     3541 accccatgca tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     3601 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     3661 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3721 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3781 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     3841 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     3901 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     3961 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4021 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     4081 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4141 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4201 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     4261 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4321 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4381 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     4441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4501 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4621 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4681 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4741 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4801 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4861 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     4921 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4981 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5041 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5101 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5161 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5221 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5281 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5341 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5401 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5461 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5521 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5581 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5641 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5701 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5761 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5821 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5881 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5941 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6001 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6661 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6721 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6781 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6841 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6901 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6961 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7021 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     7081 acaaaagctg gagctgca
//
