LOCUS       sequence-343451-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DHX58 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-DHX58-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        150..348
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     303..323
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             366..546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    593..718
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1215..1448
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1633..1677
                     /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    1975..2092
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2229..2636
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3351..3365
                     /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             3499..3679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3750..4338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3830..3849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4509..5300,5301..5369))
                     /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     5132..5151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5370..5474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5549..5568)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5742..5762)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5747..6076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5927..6062
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5989..6008
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        join(6088..6319,1..148)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
       61 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      121 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      181 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      241 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      301 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct
      361 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
      421 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      481 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      541 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
      601 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
      661 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
      721 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
      781 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
      841 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
      901 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
      961 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     1021 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct
     1081 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1141 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1201 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1261 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1321 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1381 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1441 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1501 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     1561 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     1621 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     1681 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     1741 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     1801 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     1861 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     1921 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
     1981 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2041 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     2101 ttacagggac agcagagatc cagtttggtt aattaactgc agccccgata aaataaaaga
     2161 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     2221 tagctgcagt aacgccattt tgcaaggcat ggaaaaatac caaaccaaga atagagaagt
     2281 tcagatcaag ggcgggtaca tgaaaatagc taacgttggg ccaaacagga tatctgcggt
     2341 gagcagtttc ggccccggcc cggggccaag aacagatggt caccgcagtt tcggccccgg
     2401 cccgaggcca agaacagatg gtccccagat atggcccaac cctcagcagt ttcttaagac
     2461 ccatcagatg tttccaggct cccccaagga cctgaaatga ccctgcgcct tatttgaatt
     2521 aaccaatcag cctgcttctc gcttctgttc gcgcgcttct gcttcccgag ctctataaaa
     2581 gagctcacaa cccctcactc ggcgcgccag tcctccgaca gactgagtcg cccgggggtc
     2641 tagaacgcgt gccgccatga ccgaatacaa acctaccgtg aggctggcta caagagatga
     2701 tgtcccaagg gctgtgagaa cactggccgc cgcttttgcc gattaccctg ccacacgcca
     2761 cactgtggac ccagatcggc atatcgagag agtgactgag ctgcaggaac tgttcctgac
     2821 ccgagtgggc ctggacattg ggaaggtctg ggtcgcagac gatggagcag ctgtggctgt
     2881 ctggaccaca ccagagagcg tggaagccgg agctgtcttt gcagagatcg gccctagaat
     2941 ggcagaactg agcggctcca ggctggcagc acagcagcag atggagggac tgctggcccc
     3001 acacaggcct aaggaaccag catggttcct ggctaccgtg ggggtctctc ctgaccatca
     3061 gggcaaagga ctgggaagtg ctgtggtcct gccaggagtg gaggctgcag aacgagctgg
     3121 agtccctgca tttctggaga cctctgctcc acgaaacctg cccttctatg aacggctggg
     3181 ctttactgtg accgcagatg tggaggtccc cgaaggacct aggacctggt gcatgacacg
     3241 caaacccggc gcctgagcga tcgccgcggc cgccttctta acccaacaga aggctcgaga
     3301 aggtatattg ctgttgacag tgagcgccac gcacaaggac accgtctata gtgaagccac
     3361 agatgtatag acggtgtcct tgtgcgtgtt gcctactgcc tcggacttca aggggctaga
     3421 attcggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     3481 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     3541 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     3601 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     3661 gtgtggaaaa tctctagcag ggcccgtttc atgtgagcaa aaggccagca aaaggccagg
     3721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4501 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4621 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4801 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5221 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5281 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5341 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5401 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5461 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     5521 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5581 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     5641 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     5701 ttgcgctgct tcgcgatgta cgggccagat atacgcgctg tggaatgtgt gtcagttagg
     5761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5821 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5881 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5941 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6001 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6061 cctaggcttt tgcaaaaagc tcgcgttgac attgattatt gactagttat taatagtaat
     6121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     6181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     6241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     6301 ggtaaactgc ccacttggc
//
