LOCUS       sequence-343338-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DDX17 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-DDX17-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        433..631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     586..606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             649..829
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    876..1001
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1498..1731
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1916..1960
                     /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    2258..2375
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2512..2919
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3634..3648
                     /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             3782..3962
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4033..4621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4113..4132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4792..5583,5584..5652))
                     /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     5415..5434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5653..5757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5832..5851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6025..6045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6030..6319,1..40)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6210..6319,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6272..6291
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgcgt tgacattgat
       61 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      421 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      481 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      541 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      601 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      661 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      721 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg agactctggt
      781 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      841 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      901 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      961 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1021 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1081 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1141 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1201 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1261 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1321 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
     1381 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1441 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1501 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1561 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1621 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1681 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1741 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1801 taaatctctg gaacagattt ggaatcacac gacttggatg gagtgggaca gagaaattaa
     1861 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1921 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1981 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     2041 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     2101 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     2161 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
     2221 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     2281 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2341 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     2401 atccagtttg gttaattaac tgcagccccg ataaaataaa agattttatt tagtctccag
     2461 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctgc agtaacgcca
     2521 ttttgcaagg catggaaaaa taccaaacca agaatagaga agttcagatc aagggcgggt
     2581 acatgaaaat agctaacgtt gggccaaaca ggatatctgc ggtgagcagt ttcggccccg
     2641 gcccggggcc aagaacagat ggtcaccgca gtttcggccc cggcccgagg ccaagaacag
     2701 atggtcccca gatatggccc aaccctcagc agtttcttaa gacccatcag atgtttccag
     2761 gctcccccaa ggacctgaaa tgaccctgcg ccttatttga attaaccaat cagcctgctt
     2821 ctcgcttctg ttcgcgcgct tctgcttccc gagctctata aaagagctca caacccctca
     2881 ctcggcgcgc cagtcctccg acagactgag tcgcccgggg gtctagaacg cgtgccgcca
     2941 tgaccgaata caaacctacc gtgaggctgg ctacaagaga tgatgtccca agggctgtga
     3001 gaacactggc cgccgctttt gccgattacc ctgccacacg ccacactgtg gacccagatc
     3061 ggcatatcga gagagtgact gagctgcagg aactgttcct gacccgagtg ggcctggaca
     3121 ttgggaaggt ctgggtcgca gacgatggag cagctgtggc tgtctggacc acaccagaga
     3181 gcgtggaagc cggagctgtc tttgcagaga tcggccctag aatggcagaa ctgagcggct
     3241 ccaggctggc agcacagcag cagatggagg gactgctggc cccacacagg cctaaggaac
     3301 cagcatggtt cctggctacc gtgggggtct ctcctgacca tcagggcaaa ggactgggaa
     3361 gtgctgtggt cctgccagga gtggaggctg cagaacgagc tggagtccct gcatttctgg
     3421 agacctctgc tccacgaaac ctgcccttct atgaacggct gggctttact gtgaccgcag
     3481 atgtggaggt ccccgaagga cctaggacct ggtgcatgac acgcaaaccc ggcgcctgag
     3541 cgatcgccgc ggccgccttc ttaacccaac agaaggctcg agaaggtata ttgctgttga
     3601 cagtgagcgc caatcccaat ctgatgtatc atagtgaagc cacagatgta tgatacatca
     3661 gattgggatt gttgcctact gcctcggact tcaaggggct agaattcggc agctgtagat
     3721 cttagccact ttttaaaaga aaagggggga ctggaagggc taactgcatc cggactgtac
     3781 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     3841 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     3901 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     3961 cagggcccgt ttcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5761 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     5821 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     5881 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     5941 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     6001 gtacgggcca gatatacgcg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     6061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6181 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     6301 cctctgagct attccagaa
//
