LOCUS       sequence-343341-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IFI16 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-IFI16-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        127..506
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        508..706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     661..681
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             724..904
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    951..1076
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1573..1806
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1991..2035
                     /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    2333..2450
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2587..2994
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3709..3723
                     /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             3857..4037
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4108..4696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4188..4207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4867..5658,5659..5727))
                     /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     5490..5509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5728..5832)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5907..5926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6100..6120)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6105..6319,1..115)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6285..6319,1..101)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
       61 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      121 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      181 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      241 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      301 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      361 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      421 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      481 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      541 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      601 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      661 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      721 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      781 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      841 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      901 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
      961 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1021 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1081 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1141 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1201 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1261 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1321 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1381 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
     1441 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     1501 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     1561 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     1621 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     1681 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     1741 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     1801 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     1861 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     1921 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     1981 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2041 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2101 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2161 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     2221 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     2281 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     2341 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2401 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2461 acagggacag cagagatcca gtttggttaa ttaactgcag ccccgataaa ataaaagatt
     2521 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     2581 gctgcagtaa cgccattttg caaggcatgg aaaaatacca aaccaagaat agagaagttc
     2641 agatcaaggg cgggtacatg aaaatagcta acgttgggcc aaacaggata tctgcggtga
     2701 gcagtttcgg ccccggcccg gggccaagaa cagatggtca ccgcagtttc ggccccggcc
     2761 cgaggccaag aacagatggt ccccagatat ggcccaaccc tcagcagttt cttaagaccc
     2821 atcagatgtt tccaggctcc cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa
     2881 ccaatcagcc tgcttctcgc ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga
     2941 gctcacaacc cctcactcgg cgcgccagtc ctccgacaga ctgagtcgcc cgggggtcta
     3001 gaacgcgtgc cgccatgacc gaatacaaac ctaccgtgag gctggctaca agagatgatg
     3061 tcccaagggc tgtgagaaca ctggccgccg cttttgccga ttaccctgcc acacgccaca
     3121 ctgtggaccc agatcggcat atcgagagag tgactgagct gcaggaactg ttcctgaccc
     3181 gagtgggcct ggacattggg aaggtctggg tcgcagacga tggagcagct gtggctgtct
     3241 ggaccacacc agagagcgtg gaagccggag ctgtctttgc agagatcggc cctagaatgg
     3301 cagaactgag cggctccagg ctggcagcac agcagcagat ggagggactg ctggccccac
     3361 acaggcctaa ggaaccagca tggttcctgg ctaccgtggg ggtctctcct gaccatcagg
     3421 gcaaaggact gggaagtgct gtggtcctgc caggagtgga ggctgcagaa cgagctggag
     3481 tccctgcatt tctggagacc tctgctccac gaaacctgcc cttctatgaa cggctgggct
     3541 ttactgtgac cgcagatgtg gaggtccccg aaggacctag gacctggtgc atgacacgca
     3601 aacccggcgc ctgagcgatc gccgcggccg ccttcttaac ccaacagaag gctcgagaag
     3661 gtatattgct gttgacagtg agcgccaaga acattgttct actaaatagt gaagccacag
     3721 atgtatttag tagaacaatg ttcttgttgc ctactgcctc ggacttcaag gggctagaat
     3781 tcggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaact
     3841 gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3901 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3961 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4021 gtggaaaatc tctagcaggg cccgtttcat gtgagcaaaa ggccagcaaa aggccaggaa
     4081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4681 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4801 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4861 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4921 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4981 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5041 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5101 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5161 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5221 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5281 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5341 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5401 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5461 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5521 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5581 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5641 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5701 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5761 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5821 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     5881 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5941 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     6001 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     6061 gcgctgcttc gcgatgtacg ggccagatat acgcgctgtg gaatgtgtgt cagttagggt
     6121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6181 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6241 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6301 cgcccagttc cgcccattc
//
