LOCUS       sequence-343175-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IFIH1 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-IFIH1-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        340..538
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     493..513
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             556..736
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    783..908
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1405..1638
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1823..1867
                     /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    2165..2282
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2419..2826
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3541..3555
                     /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             3689..3869
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3940..4528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4020..4039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4699..5490,5491..5559))
                     /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     5322..5341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5560..5664)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5739..5758)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5932..5952)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5937..6266
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6117..6252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6179..6198
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        join(6278..6319,1..338)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
       61 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      121 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      181 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      241 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      301 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      361 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      421 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      481 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg
      541 cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      721 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
      781 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
      841 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
      901 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
      961 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     1021 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1081 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1141 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1201 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     1261 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1321 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1381 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1441 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1501 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1561 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1621 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1681 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1741 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     1801 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     1861 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     1921 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     1981 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2041 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2101 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     2161 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2221 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2281 ttcgggttta ttacagggac agcagagatc cagtttggtt aattaactgc agccccgata
     2341 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     2401 tttggcaagc tagctgcagt aacgccattt tgcaaggcat ggaaaaatac caaaccaaga
     2461 atagagaagt tcagatcaag ggcgggtaca tgaaaatagc taacgttggg ccaaacagga
     2521 tatctgcggt gagcagtttc ggccccggcc cggggccaag aacagatggt caccgcagtt
     2581 tcggccccgg cccgaggcca agaacagatg gtccccagat atggcccaac cctcagcagt
     2641 ttcttaagac ccatcagatg tttccaggct cccccaagga cctgaaatga ccctgcgcct
     2701 tatttgaatt aaccaatcag cctgcttctc gcttctgttc gcgcgcttct gcttcccgag
     2761 ctctataaaa gagctcacaa cccctcactc ggcgcgccag tcctccgaca gactgagtcg
     2821 cccgggggtc tagaacgcgt gccgccatga ccgaatacaa acctaccgtg aggctggcta
     2881 caagagatga tgtcccaagg gctgtgagaa cactggccgc cgcttttgcc gattaccctg
     2941 ccacacgcca cactgtggac ccagatcggc atatcgagag agtgactgag ctgcaggaac
     3001 tgttcctgac ccgagtgggc ctggacattg ggaaggtctg ggtcgcagac gatggagcag
     3061 ctgtggctgt ctggaccaca ccagagagcg tggaagccgg agctgtcttt gcagagatcg
     3121 gccctagaat ggcagaactg agcggctcca ggctggcagc acagcagcag atggagggac
     3181 tgctggcccc acacaggcct aaggaaccag catggttcct ggctaccgtg ggggtctctc
     3241 ctgaccatca gggcaaagga ctgggaagtg ctgtggtcct gccaggagtg gaggctgcag
     3301 aacgagctgg agtccctgca tttctggaga cctctgctcc acgaaacctg cccttctatg
     3361 aacggctggg ctttactgtg accgcagatg tggaggtccc cgaaggacct aggacctggt
     3421 gcatgacacg caaacccggc gcctgagcga tcgccgcggc cgccttctta acccaacaga
     3481 aggctcgaga aggtatattg ctgttgacag tgagcgacga gagaagatga tgtataaata
     3541 gtgaagccac agatgtattt atacatcatc ttctctcggt gcctactgcc tcggacttca
     3601 aggggctaga attcggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     3661 gaagggctaa ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     3721 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3781 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3841 cttttagtca gtgtggaaaa tctctagcag ggcccgtttc atgtgagcaa aaggccagca
     3901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4381 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4621 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4681 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4741 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4801 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4861 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4921 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4981 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5041 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5101 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5161 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5221 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5281 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5341 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5401 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5461 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5521 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5581 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5641 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     5701 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     5761 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     5821 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     5881 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgctg tggaatgtgt
     5941 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6001 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     6061 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6121 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6181 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     6241 tttttggagg cctaggcttt tgcaaaaagc tcgcgttgac attgattatt gactagttat
     6301 taatagtaat caattacgg
//
