LOCUS       sequence-343458-        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-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        429..627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     582..602
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             645..825
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    872..997
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1494..1727
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1912..1956
                     /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    2254..2371
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2508..2915
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3630..3644
                     /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             3778..3958
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4029..4617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4109..4128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4788..5579,5580..5648))
                     /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     5411..5430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5649..5753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5828..5847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6021..6041)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6026..6319,1..36)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6206..6319,1..22)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6268..6287
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcgcgttgac attgattatt
       61 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      601 tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag
      661 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
      721 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
      781 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
      841 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
      901 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
      961 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     1021 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     1081 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     1141 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     1201 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     1261 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     1321 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1381 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1441 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1501 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1561 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1621 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1681 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1741 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1801 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1861 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1921 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1981 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2041 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     2101 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     2161 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     2221 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     2281 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     2341 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     2401 agtttggtta attaactgca gccccgataa aataaaagat tttatttagt ctccagaaaa
     2461 aggggggaat gaaagacccc acctgtaggt ttggcaagct agctgcagta acgccatttt
     2521 gcaaggcatg gaaaaatacc aaaccaagaa tagagaagtt cagatcaagg gcgggtacat
     2581 gaaaatagct aacgttgggc caaacaggat atctgcggtg agcagtttcg gccccggccc
     2641 ggggccaaga acagatggtc accgcagttt cggccccggc ccgaggccaa gaacagatgg
     2701 tccccagata tggcccaacc ctcagcagtt tcttaagacc catcagatgt ttccaggctc
     2761 ccccaaggac ctgaaatgac cctgcgcctt atttgaatta accaatcagc ctgcttctcg
     2821 cttctgttcg cgcgcttctg cttcccgagc tctataaaag agctcacaac ccctcactcg
     2881 gcgcgccagt cctccgacag actgagtcgc ccgggggtct agaacgcgtg ccgccatgac
     2941 cgaatacaaa cctaccgtga ggctggctac aagagatgat gtcccaaggg ctgtgagaac
     3001 actggccgcc gcttttgccg attaccctgc cacacgccac actgtggacc cagatcggca
     3061 tatcgagaga gtgactgagc tgcaggaact gttcctgacc cgagtgggcc tggacattgg
     3121 gaaggtctgg gtcgcagacg atggagcagc tgtggctgtc tggaccacac cagagagcgt
     3181 ggaagccgga gctgtctttg cagagatcgg ccctagaatg gcagaactga gcggctccag
     3241 gctggcagca cagcagcaga tggagggact gctggcccca cacaggccta aggaaccagc
     3301 atggttcctg gctaccgtgg gggtctctcc tgaccatcag ggcaaaggac tgggaagtgc
     3361 tgtggtcctg ccaggagtgg aggctgcaga acgagctgga gtccctgcat ttctggagac
     3421 ctctgctcca cgaaacctgc ccttctatga acggctgggc tttactgtga ccgcagatgt
     3481 ggaggtcccc gaaggaccta ggacctggtg catgacacgc aaacccggcg cctgagcgat
     3541 cgccgcggcc gccttcttaa cccaacagaa ggctcgagaa ggtatattgc tgttgacagt
     3601 gagcgcgagc cagtacctag aacttaatag tgaagccaca gatgtattaa gttctaggta
     3661 ctggctcatg cctactgcct cggacttcaa ggggctagaa ttcggcagct gtagatctta
     3721 gccacttttt aaaagaaaag gggggactgg aagggctaac tgcatccgga ctgtactggg
     3781 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     3841 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     3901 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg
     3961 gcccgtttca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4021 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4081 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4201 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4441 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4921 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5161 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5761 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     5821 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     5881 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     5941 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     6001 gggccagata tacgcgctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     6061 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     6121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6181 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6241 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     6301 tgagctattc cagaagtag
//
