LOCUS       sequence-343773-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAVS 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-MAVS-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        94..423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      274..409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     336..355
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        435..814
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        816..1014
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     969..989
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1032..1212
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1259..1384
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1881..2114
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2299..2343
                     /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    2641..2758
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2895..3302
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           4017..4031
                     /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             4165..4345
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4416..5004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4496..4515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5175..5966,5967..6035))
                     /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     5798..5817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6036..6140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6215..6234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 cgatgtacgg gccagatata cgcgctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      361 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      421 aaaaagctcg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      481 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      541 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      601 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      661 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      721 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      781 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      841 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      901 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      961 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
     1021 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     1081 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     1141 cccgtctgtt gtgagactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     1201 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     1261 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     1321 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     1381 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     1441 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     1501 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     1561 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     1621 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     1681 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc gctgatcttc
     1741 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     1801 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     1861 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     1921 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     1981 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     2041 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     2101 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     2161 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacttgg
     2221 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     2281 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     2341 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     2401 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     2461 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     2521 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     2581 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
     2641 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     2701 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     2761 ggtttattac agggacagca gagatccagt ttggttaatt aactgcagcc ccgataaaat
     2821 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     2881 gcaagctagc tgcagtaacg ccattttgca aggcatggaa aaataccaaa ccaagaatag
     2941 agaagttcag atcaagggcg ggtacatgaa aatagctaac gttgggccaa acaggatatc
     3001 tgcggtgagc agtttcggcc ccggcccggg gccaagaaca gatggtcacc gcagtttcgg
     3061 ccccggcccg aggccaagaa cagatggtcc ccagatatgg cccaaccctc agcagtttct
     3121 taagacccat cagatgtttc caggctcccc caaggacctg aaatgaccct gcgccttatt
     3181 tgaattaacc aatcagcctg cttctcgctt ctgttcgcgc gcttctgctt cccgagctct
     3241 ataaaagagc tcacaacccc tcactcggcg cgccagtcct ccgacagact gagtcgcccg
     3301 ggggtctaga acgcgtgccg ccatgaccga atacaaacct accgtgaggc tggctacaag
     3361 agatgatgtc ccaagggctg tgagaacact ggccgccgct tttgccgatt accctgccac
     3421 acgccacact gtggacccag atcggcatat cgagagagtg actgagctgc aggaactgtt
     3481 cctgacccga gtgggcctgg acattgggaa ggtctgggtc gcagacgatg gagcagctgt
     3541 ggctgtctgg accacaccag agagcgtgga agccggagct gtctttgcag agatcggccc
     3601 tagaatggca gaactgagcg gctccaggct ggcagcacag cagcagatgg agggactgct
     3661 ggccccacac aggcctaagg aaccagcatg gttcctggct accgtggggg tctctcctga
     3721 ccatcagggc aaaggactgg gaagtgctgt ggtcctgcca ggagtggagg ctgcagaacg
     3781 agctggagtc cctgcatttc tggagacctc tgctccacga aacctgccct tctatgaacg
     3841 gctgggcttt actgtgaccg cagatgtgga ggtccccgaa ggacctagga cctggtgcat
     3901 gacacgcaaa cccggcgcct gagcgatcgc cgcggccgcc ttcttaaccc aacagaaggc
     3961 tcgagaaggt atattgctgt tgacagtgag cgaaccaatc cagcaccatc caaatagtga
     4021 agccacagat gtatttggat ggtgctggat tggtgtgcct actgcctcgg acttcaaggg
     4081 gctagaattc ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4141 ggctaactgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
     4201 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4261 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4321 tagtcagtgt ggaaaatctc tagcagggcc cgtttcatgt gagcaaaagg ccagcaaaag
     4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6181 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6241 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     6301 tttaagctac aacaaggca
//
