LOCUS       sequence-343465-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DDX3X 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-DDX3X-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        460..658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     613..633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             676..856
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    903..1028
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1525..1758
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1943..1987
                     /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    2285..2402
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2539..2946
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3661..3675
                     /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             3809..3989
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4819..5610,5611..5679))
                     /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     5442..5461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5680..5784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5859..5878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6052..6072)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6057..6319,1..67)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6237..6319,1..53)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6299..6318
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
       61 ttgcaaaaag ctcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg
      661 cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      721 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      781 tgtgcccgtc tgttgtgaga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      841 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
      901 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
      961 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     1021 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     1081 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     1141 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1201 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1261 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1321 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     1381 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1441 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1501 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1561 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1621 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1681 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1741 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1801 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1861 ttggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     1921 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     1981 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2041 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2101 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2161 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2221 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     2281 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2341 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2401 ttcgggttta ttacagggac agcagagatc cagtttggtt aattaactgc agccccgata
     2461 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     2521 tttggcaagc tagctgcagt aacgccattt tgcaaggcat ggaaaaatac caaaccaaga
     2581 atagagaagt tcagatcaag ggcgggtaca tgaaaatagc taacgttggg ccaaacagga
     2641 tatctgcggt gagcagtttc ggccccggcc cggggccaag aacagatggt caccgcagtt
     2701 tcggccccgg cccgaggcca agaacagatg gtccccagat atggcccaac cctcagcagt
     2761 ttcttaagac ccatcagatg tttccaggct cccccaagga cctgaaatga ccctgcgcct
     2821 tatttgaatt aaccaatcag cctgcttctc gcttctgttc gcgcgcttct gcttcccgag
     2881 ctctataaaa gagctcacaa cccctcactc ggcgcgccag tcctccgaca gactgagtcg
     2941 cccgggggtc tagaacgcgt gccgccatga ccgaatacaa acctaccgtg aggctggcta
     3001 caagagatga tgtcccaagg gctgtgagaa cactggccgc cgcttttgcc gattaccctg
     3061 ccacacgcca cactgtggac ccagatcggc atatcgagag agtgactgag ctgcaggaac
     3121 tgttcctgac ccgagtgggc ctggacattg ggaaggtctg ggtcgcagac gatggagcag
     3181 ctgtggctgt ctggaccaca ccagagagcg tggaagccgg agctgtcttt gcagagatcg
     3241 gccctagaat ggcagaactg agcggctcca ggctggcagc acagcagcag atggagggac
     3301 tgctggcccc acacaggcct aaggaaccag catggttcct ggctaccgtg ggggtctctc
     3361 ctgaccatca gggcaaagga ctgggaagtg ctgtggtcct gccaggagtg gaggctgcag
     3421 aacgagctgg agtccctgca tttctggaga cctctgctcc acgaaacctg cccttctatg
     3481 aacggctggg ctttactgtg accgcagatg tggaggtccc cgaaggacct aggacctggt
     3541 gcatgacacg caaacccggc gcctgagcga tcgccgcggc cgccttctta acccaacaga
     3601 aggctcgaga aggtatattg ctgttgacag tgagcgagcg ctatattcct cctcattata
     3661 gtgaagccac agatgtataa tgaggaggaa tatagcgcct gcctactgcc tcggacttca
     3721 aggggctaga attcggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     3781 gaagggctaa ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     3841 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3901 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3961 cttttagtca gtgtggaaaa tctctagcag ggcccgtttc atgtgagcaa aaggccagca
     4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     5821 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     5881 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     5941 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     6001 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgctg tggaatgtgt
     6061 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6121 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     6181 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6241 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6301 tttatgcaga ggccgaggc
//
