LOCUS       sequence-251847-        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"
     primer_bind     16..33
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    150..171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(186..191,192..209,210..216)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    224..240
                     /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     229..251
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     248..264
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     248..264
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     283..303
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        285..303
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        331..557
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             558..738
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    785..910
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1403..1636
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1821..1865
                     /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        2163..2411
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2163..2183
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2334..2353
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     ncRNA           2438..2452
                     /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    2522..2639
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2688..3198
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3117..3135
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3220..3819
                     /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(3220..3239)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3716..3736
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3947..4180
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4252..4373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4289..4308)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4343..4362
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4413..4548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4568..4587)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4569..4587)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4597..4614)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4597..4613)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4606..4628)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4755..5210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4842..4861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5052..5073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5236..5340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5341..5409,5410..6201)
                     /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(5559..5578)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6372..6960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6861..6880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
       61 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      121 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      181 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      241 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      301 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
      361 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
      421 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
      481 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
      541 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      601 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      661 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      721 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      781 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      841 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
      901 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
      961 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     1021 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1081 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1141 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1201 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     1261 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     1321 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     1381 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     1441 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     1501 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1561 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1621 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1681 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1741 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1801 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     1861 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     1921 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     1981 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     2041 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     2101 tagtgaacgg atctcgacgg tatcgatcac gagactagcc tcgagcggcc gcccccttca
     2161 ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     2221 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     2281 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     2341 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     2401 gacgaaacac cggtacccag gaagacattc tggatctgtg aagccacaga tgggatccag
     2461 aatgtcttcc tgggtatttt taattctcga cctcgagaca aatggcagta ttcatccaca
     2521 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     2581 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     2641 gggtttatta cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg
     2701 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     2761 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     2821 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     2881 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     2941 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     3001 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     3061 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     3121 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     3181 accgacctct ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg
     3241 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3301 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3361 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3421 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3481 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3541 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3601 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3661 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3721 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     3781 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     3841 cgcccgaccg aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac
     3901 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     3961 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     4021 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     4081 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     4141 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     4201 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     4261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4381 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4441 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4501 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     4561 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     4621 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4681 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4741 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4801 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4861 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4921 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4981 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5041 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5101 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5161 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     5221 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5281 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5341 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5401 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5461 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5521 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5581 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5641 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5701 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5761 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5821 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5881 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5941 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6001 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6061 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6121 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6181 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6241 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6301 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6361 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6421 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6481 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6541 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6601 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6661 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6721 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6781 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6841 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6901 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6961 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7021 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7081 gataccgctc gccgcagc
//
