LOCUS       sequence-343163-        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-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        31..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             628..808
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    855..980
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1477..1710
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1895..1939
                     /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    2237..2354
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2491..2898
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3613..3627
                     /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             3761..3941
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4012..4600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4092..4111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4771..5562,5563..5631))
                     /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     5394..5413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5632..5736)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5811..5830)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6004..6024)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6009..6319,1..19)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6189..6319,1..5)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6251..6270
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aggcctaggc ttttgcaaaa agctcgcgtt gacattgatt attgactagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
      661 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      721 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      781 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      841 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      901 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      961 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1021 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1081 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1141 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1201 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1261 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1321 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1381 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1441 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1501 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1561 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1621 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1681 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1741 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1801 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1861 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1921 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1981 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     2041 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     2101 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     2161 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg
     2221 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     2281 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     2341 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaact
     2401 gcagccccga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     2461 cccacctgta ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat
     2521 accaaaccaa gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg
     2581 ggccaaacag gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg
     2641 gtcaccgcag tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca
     2701 accctcagca gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat
     2761 gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt
     2821 ctgcttcccg agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga
     2881 cagactgagt cgcccggggg tctagaacgc gtgccgccat gaccgaatac aaacctaccg
     2941 tgaggctggc tacaagagat gatgtcccaa gggctgtgag aacactggcc gccgcttttg
     3001 ccgattaccc tgccacacgc cacactgtgg acccagatcg gcatatcgag agagtgactg
     3061 agctgcagga actgttcctg acccgagtgg gcctggacat tgggaaggtc tgggtcgcag
     3121 acgatggagc agctgtggct gtctggacca caccagagag cgtggaagcc ggagctgtct
     3181 ttgcagagat cggccctaga atggcagaac tgagcggctc caggctggca gcacagcagc
     3241 agatggaggg actgctggcc ccacacaggc ctaaggaacc agcatggttc ctggctaccg
     3301 tgggggtctc tcctgaccat cagggcaaag gactgggaag tgctgtggtc ctgccaggag
     3361 tggaggctgc agaacgagct ggagtccctg catttctgga gacctctgct ccacgaaacc
     3421 tgcccttcta tgaacggctg ggctttactg tgaccgcaga tgtggaggtc cccgaaggac
     3481 ctaggacctg gtgcatgaca cgcaaacccg gcgcctgagc gatcgccgcg gccgccttct
     3541 taacccaaca gaaggctcga gaaggtatat tgctgttgac agtgagcgcg gatattaaag
     3601 aatgtaacat tagtgaagcc acagatgtaa tgttacattc tttaatatcc atgcctactg
     3661 cctcggactt caaggggcta gaattcggca gctgtagatc ttagccactt tttaaaagaa
     3721 aaggggggac tggaagggct aactgcatcc ggactgtact gggtctctct ggttagacca
     3781 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     3841 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     3901 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt tcatgtgagc
     3961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4441 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4681 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4741 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4801 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4861 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4921 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4981 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5041 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5101 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5161 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5221 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5281 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5341 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5401 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5461 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5521 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5581 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5761 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5821 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5881 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     5941 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
     6001 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     6061 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6241 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     6301 agtgaggagg cttttttgg
//
