LOCUS       sequence-343774-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ILF2 knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6319)
  AUTHORS   Guney MH, Nagalekshmi K, McCauley SM, Carbone C, Aydemir O, Luban J
  TITLE     IFIH1 (MDA5) is required for innate immune detection of 
            intron-containing RNA expressed from the HIV-1 provirus.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Jul 16;121(29):e2404349121. doi: 
            10.1073/pnas.2404349121. Epub 2024 Jul 10.
  PUBMED    38985764
REFERENCE   2  (bases 1 to 6319)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-07-16"; volume: "121(29):e2404349121. 
            doi"; pages: " 10.1073/pnas.2404349121. Epub 2024 Jul 10"
COMMENT     Sequence Label: pAPM-miR30-ILF2-ts1
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..45
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             88..268
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    315..440
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    937..1170
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1355..1399
                     /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"
     misc_feature    1697..1814
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1951..2358
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3073..3087
                     /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"
     LTR             3221..3401
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3472..4060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3552..3571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4231..5022,5023..5091))
                     /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     4854..4873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5092..5196)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5271..5290)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5464..5484)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5469..5798
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5649..5784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5711..5730
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5810..6189
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6191..6319,1..70)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
       61 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
      121 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      181 cttcaagtag tgtgtgcccg tctgttgtga gactctggta actagagatc cctcagaccc
      241 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      301 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      361 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      421 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
      481 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
      541 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
      601 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
      661 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
      721 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
      781 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
      841 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
      901 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
      961 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1021 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1081 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1141 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1201 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1261 gaatcacacg acttggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1321 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1381 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1441 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     1501 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     1561 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     1621 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg
     1681 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     1741 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     1801 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaact
     1861 gcagccccga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     1921 cccacctgta ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat
     1981 accaaaccaa gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg
     2041 ggccaaacag gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg
     2101 gtcaccgcag tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca
     2161 accctcagca gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat
     2221 gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt
     2281 ctgcttcccg agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga
     2341 cagactgagt cgcccggggg tctagaacgc gtgccgccat gaccgaatac aaacctaccg
     2401 tgaggctggc tacaagagat gatgtcccaa gggctgtgag aacactggcc gccgcttttg
     2461 ccgattaccc tgccacacgc cacactgtgg acccagatcg gcatatcgag agagtgactg
     2521 agctgcagga actgttcctg acccgagtgg gcctggacat tgggaaggtc tgggtcgcag
     2581 acgatggagc agctgtggct gtctggacca caccagagag cgtggaagcc ggagctgtct
     2641 ttgcagagat cggccctaga atggcagaac tgagcggctc caggctggca gcacagcagc
     2701 agatggaggg actgctggcc ccacacaggc ctaaggaacc agcatggttc ctggctaccg
     2761 tgggggtctc tcctgaccat cagggcaaag gactgggaag tgctgtggtc ctgccaggag
     2821 tggaggctgc agaacgagct ggagtccctg catttctgga gacctctgct ccacgaaacc
     2881 tgcccttcta tgaacggctg ggctttactg tgaccgcaga tgtggaggtc cccgaaggac
     2941 ctaggacctg gtgcatgaca cgcaaacccg gcgcctgagc gatcgccgcg gccgccttct
     3001 taacccaaca gaaggctcga gaaggtatat tgctgttgac agtgagcgcc aggatccttc
     3061 tgaagtttta tagtgaagcc acagatgtat aaaacttcag aaggatcctg ttgcctactg
     3121 cctcggactt caaggggcta gaattcggca gctgtagatc ttagccactt tttaaaagaa
     3181 aaggggggac tggaagggct aactgcatcc ggactgtact gggtctctct ggttagacca
     3241 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     3301 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     3361 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt tcatgtgagc
     3421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5221 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5281 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5341 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     5401 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
     5461 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5521 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5641 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5701 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5761 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg acattgatta
     5821 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5881 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5941 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     6001 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     6061 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     6121 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6181 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     6241 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     6301 caacgggact ttccaaaat
//
