LOCUS       sequence-343466-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ILF3 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-ILF3-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     559..579
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             622..802
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    849..974
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1471..1704
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1889..1933
                     /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    2231..2348
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2485..2892
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3607..3621
                     /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             3755..3935
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4006..4594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4086..4105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4765..5556,5557..5625))
                     /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     5388..5407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5626..5730)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5805..5824)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5998..6018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6003..6319,1..13)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6183..6318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6245..6264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aggcttttgc aaaaagctcg cgttgacatt gattattgac tagttattaa tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      421 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      481 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      541 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      601 gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
      661 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
      721 gtagtgtgtg cccgtctgtt gtgagactct ggtaactaga gatccctcag acccttttag
      781 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
      841 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
      901 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
      961 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     1021 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     1081 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     1141 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     1201 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     1261 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc
     1321 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     1381 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     1441 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     1501 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     1561 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     1621 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     1681 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     1741 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     1801 cacgacttgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     1861 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     1921 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     1981 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     2041 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     2101 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     2161 cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat
     2221 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     2281 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     2341 aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt aactgcagcc
     2401 ccgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     2461 tgtaggtttg gcaagctagc tgcagtaacg ccattttgca aggcatggaa aaataccaaa
     2521 ccaagaatag agaagttcag atcaagggcg ggtacatgaa aatagctaac gttgggccaa
     2581 acaggatatc tgcggtgagc agtttcggcc ccggcccggg gccaagaaca gatggtcacc
     2641 gcagtttcgg ccccggcccg aggccaagaa cagatggtcc ccagatatgg cccaaccctc
     2701 agcagtttct taagacccat cagatgtttc caggctcccc caaggacctg aaatgaccct
     2761 gcgccttatt tgaattaacc aatcagcctg cttctcgctt ctgttcgcgc gcttctgctt
     2821 cccgagctct ataaaagagc tcacaacccc tcactcggcg cgccagtcct ccgacagact
     2881 gagtcgcccg ggggtctaga acgcgtgccg ccatgaccga atacaaacct accgtgaggc
     2941 tggctacaag agatgatgtc ccaagggctg tgagaacact ggccgccgct tttgccgatt
     3001 accctgccac acgccacact gtggacccag atcggcatat cgagagagtg actgagctgc
     3061 aggaactgtt cctgacccga gtgggcctgg acattgggaa ggtctgggtc gcagacgatg
     3121 gagcagctgt ggctgtctgg accacaccag agagcgtgga agccggagct gtctttgcag
     3181 agatcggccc tagaatggca gaactgagcg gctccaggct ggcagcacag cagcagatgg
     3241 agggactgct ggccccacac aggcctaagg aaccagcatg gttcctggct accgtggggg
     3301 tctctcctga ccatcagggc aaaggactgg gaagtgctgt ggtcctgcca ggagtggagg
     3361 ctgcagaacg agctggagtc cctgcatttc tggagacctc tgctccacga aacctgccct
     3421 tctatgaacg gctgggcttt actgtgaccg cagatgtgga ggtccccgaa ggacctagga
     3481 cctggtgcat gacacgcaaa cccggcgcct gagcgatcgc cgcggccgcc ttcttaaccc
     3541 aacagaaggc tcgagaaggt atattgctgt tgacagtgag cgaccagagg acgacagtaa
     3601 agaatagtga agccacagat gtattcttta ctgtcgtcct ctgggtgcct actgcctcgg
     3661 acttcaaggg gctagaattc ggcagctgta gatcttagcc actttttaaa agaaaagggg
     3721 ggactggaag ggctaactgc atccggactg tactgggtct ctctggttag accagatctg
     3781 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3841 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3901 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttcatgt gagcaaaagg
     3961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4681 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4741 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4801 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4861 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4921 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4981 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5041 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5101 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5161 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5221 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5281 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5341 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5401 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5461 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5521 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5581 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5641 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5701 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5761 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     5821 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     5881 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     5941 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgctgtgga
     6001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6241 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6301 gaggcttttt tggaggcct
//
