LOCUS       sequence-343346-        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-ts1
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..98)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        83..412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      263..398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     325..344
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        424..803
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        805..1003
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     958..978
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1021..1201
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1248..1373
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1870..2103
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2288..2332
                     /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    2630..2747
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2884..3291
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           4006..4020
                     /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             4154..4334
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4405..4993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4485..4504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5164..5955,5956..6024))
                     /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     5787..5806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6025..6129)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6204..6223)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
       61 ccagatatac gcgctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
      121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
      181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
      241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
      301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
      361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcgc
      421 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      481 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      541 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      601 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      661 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      721 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      781 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      841 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      901 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      961 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
     1021 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     1081 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     1141 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     1201 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     1261 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     1321 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     1381 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     1441 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     1501 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     1561 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     1621 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     1681 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     1741 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     1801 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     1861 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     1921 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     1981 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     2041 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     2101 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     2161 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     2221 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     2281 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     2341 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     2401 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     2461 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     2521 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     2581 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     2641 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     2701 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     2761 gggacagcag agatccagtt tggttaatta actgcagccc cgataaaata aaagatttta
     2821 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     2881 gcagtaacgc cattttgcaa ggcatggaaa aataccaaac caagaataga gaagttcaga
     2941 tcaagggcgg gtacatgaaa atagctaacg ttgggccaaa caggatatct gcggtgagca
     3001 gtttcggccc cggcccgggg ccaagaacag atggtcaccg cagtttcggc cccggcccga
     3061 ggccaagaac agatggtccc cagatatggc ccaaccctca gcagtttctt aagacccatc
     3121 agatgtttcc aggctccccc aaggacctga aatgaccctg cgccttattt gaattaacca
     3181 atcagcctgc ttctcgcttc tgttcgcgcg cttctgcttc ccgagctcta taaaagagct
     3241 cacaacccct cactcggcgc gccagtcctc cgacagactg agtcgcccgg gggtctagaa
     3301 cgcgtgccgc catgaccgaa tacaaaccta ccgtgaggct ggctacaaga gatgatgtcc
     3361 caagggctgt gagaacactg gccgccgctt ttgccgatta ccctgccaca cgccacactg
     3421 tggacccaga tcggcatatc gagagagtga ctgagctgca ggaactgttc ctgacccgag
     3481 tgggcctgga cattgggaag gtctgggtcg cagacgatgg agcagctgtg gctgtctgga
     3541 ccacaccaga gagcgtggaa gccggagctg tctttgcaga gatcggccct agaatggcag
     3601 aactgagcgg ctccaggctg gcagcacagc agcagatgga gggactgctg gccccacaca
     3661 ggcctaagga accagcatgg ttcctggcta ccgtgggggt ctctcctgac catcagggca
     3721 aaggactggg aagtgctgtg gtcctgccag gagtggaggc tgcagaacga gctggagtcc
     3781 ctgcatttct ggagacctct gctccacgaa acctgccctt ctatgaacgg ctgggcttta
     3841 ctgtgaccgc agatgtggag gtccccgaag gacctaggac ctggtgcatg acacgcaaac
     3901 ccggcgcctg agcgatcgcc gcggccgcct tcttaaccca acagaaggct cgagaaggta
     3961 tattgctgtt gacagtgagc gacagcttga aggagaaatt caatagtgaa gccacagatg
     4021 tattgaattt ctccttcaag ctggtgccta ctgcctcgga cttcaagggg ctagaattcg
     4081 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaactgca
     4141 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4201 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4261 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4321 gaaaatctct agcagggccc gtttcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4981 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5041 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5101 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5161 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5221 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5341 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5581 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5821 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6001 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6061 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6121 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6181 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6241 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     6301 acaaggcaag gcttgaccg
//
