LOCUS       sequence-343174-        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-ts1
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..61
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        141..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        522..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             738..918
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    965..1090
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1587..1820
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2005..2049
                     /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    2347..2464
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2601..3008
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3723..3737
                     /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             3871..4051
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4122..4710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4202..4221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4881..5672,5673..5741))
                     /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     5504..5523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5742..5846)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5921..5940)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6114..6134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6119..6319,1..129)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6299..6319,1..115)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
       61 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      121 ttttgcaaaa agctcgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      781 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      841 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      901 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      961 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     1021 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     1081 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     1141 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     1201 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1261 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1321 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1381 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg
     1441 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     1501 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     1561 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     1621 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     1681 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     1741 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     1801 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     1861 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     1921 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     1981 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     2041 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     2101 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     2161 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     2221 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     2281 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat
     2341 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2401 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2461 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaact gcagccccga
     2521 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     2581 ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat accaaaccaa
     2641 gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg ggccaaacag
     2701 gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg gtcaccgcag
     2761 tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca accctcagca
     2821 gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat gaccctgcgc
     2881 cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg
     2941 agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga cagactgagt
     3001 cgcccggggg tctagaacgc gtgccgccat gaccgaatac aaacctaccg tgaggctggc
     3061 tacaagagat gatgtcccaa gggctgtgag aacactggcc gccgcttttg ccgattaccc
     3121 tgccacacgc cacactgtgg acccagatcg gcatatcgag agagtgactg agctgcagga
     3181 actgttcctg acccgagtgg gcctggacat tgggaaggtc tgggtcgcag acgatggagc
     3241 agctgtggct gtctggacca caccagagag cgtggaagcc ggagctgtct ttgcagagat
     3301 cggccctaga atggcagaac tgagcggctc caggctggca gcacagcagc agatggaggg
     3361 actgctggcc ccacacaggc ctaaggaacc agcatggttc ctggctaccg tgggggtctc
     3421 tcctgaccat cagggcaaag gactgggaag tgctgtggtc ctgccaggag tggaggctgc
     3481 agaacgagct ggagtccctg catttctgga gacctctgct ccacgaaacc tgcccttcta
     3541 tgaacggctg ggctttactg tgaccgcaga tgtggaggtc cccgaaggac ctaggacctg
     3601 gtgcatgaca cgcaaacccg gcgcctgagc gatcgccgcg gccgccttct taacccaaca
     3661 gaaggctcga gaaggtatat tgctgttgac agtgagcgac ctaccaaggc cacttctatt
     3721 tagtgaagcc acagatgtaa atagaagtgg ccttggtagg gtgcctactg cctcggactt
     3781 caaggggcta gaattcggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     3841 tggaagggct aactgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
     3901 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3961 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     4021 cccttttagt cagtgtggaa aatctctagc agggcccgtt tcatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     5881 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     5941 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     6001 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6061 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc tgtggaatgt
     6121 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6181 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6241 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6301 cccgccccta actccgccc
//
