LOCUS       sequence-343450-        6319 bp    DNA     circular SYN 15-APR-2025
DEFINITION  IFI16 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 Apr 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-IFI16-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        443..641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     596..616
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             659..839
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    886..1011
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1508..1741
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1926..1970
                     /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    2268..2385
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2522..2929
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3644..3658
                     /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             3792..3972
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4043..4631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4123..4142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4802..5593,5594..5662))
                     /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     5425..5444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5663..5767)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5842..5861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6035..6055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6040..6319,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6220..6319,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6282..6301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgcgt
       61 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      121 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      181 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      481 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      541 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      601 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
      661 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      721 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      781 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      841 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
      901 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
      961 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     1021 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     1081 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     1141 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     1201 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     1261 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     1321 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag
     1381 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1441 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1501 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     1561 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     1621 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     1681 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     1741 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     1801 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     1861 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     1921 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     1981 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     2041 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     2101 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     2161 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     2221 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     2281 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2341 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     2401 gacagcagag atccagtttg gttaattaac tgcagccccg ataaaataaa agattttatt
     2461 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctgc
     2521 agtaacgcca ttttgcaagg catggaaaaa taccaaacca agaatagaga agttcagatc
     2581 aagggcgggt acatgaaaat agctaacgtt gggccaaaca ggatatctgc ggtgagcagt
     2641 ttcggccccg gcccggggcc aagaacagat ggtcaccgca gtttcggccc cggcccgagg
     2701 ccaagaacag atggtcccca gatatggccc aaccctcagc agtttcttaa gacccatcag
     2761 atgtttccag gctcccccaa ggacctgaaa tgaccctgcg ccttatttga attaaccaat
     2821 cagcctgctt ctcgcttctg ttcgcgcgct tctgcttccc gagctctata aaagagctca
     2881 caacccctca ctcggcgcgc cagtcctccg acagactgag tcgcccgggg gtctagaacg
     2941 cgtgccgcca tgaccgaata caaacctacc gtgaggctgg ctacaagaga tgatgtccca
     3001 agggctgtga gaacactggc cgccgctttt gccgattacc ctgccacacg ccacactgtg
     3061 gacccagatc ggcatatcga gagagtgact gagctgcagg aactgttcct gacccgagtg
     3121 ggcctggaca ttgggaaggt ctgggtcgca gacgatggag cagctgtggc tgtctggacc
     3181 acaccagaga gcgtggaagc cggagctgtc tttgcagaga tcggccctag aatggcagaa
     3241 ctgagcggct ccaggctggc agcacagcag cagatggagg gactgctggc cccacacagg
     3301 cctaaggaac cagcatggtt cctggctacc gtgggggtct ctcctgacca tcagggcaaa
     3361 ggactgggaa gtgctgtggt cctgccagga gtggaggctg cagaacgagc tggagtccct
     3421 gcatttctgg agacctctgc tccacgaaac ctgcccttct atgaacggct gggctttact
     3481 gtgaccgcag atgtggaggt ccccgaagga cctaggacct ggtgcatgac acgcaaaccc
     3541 ggcgcctgag cgatcgccgc ggccgccttc ttaacccaac agaaggctcg agaaggtata
     3601 ttgctgttga cagtgagcgc tacaagaaca ttgttctact atagtgaagc cacagatgta
     3661 tagtagaaca atgttcttgt attgcctact gcctcggact tcaaggggct agaattcggc
     3721 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taactgcatc
     3781 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     3841 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     3901 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     3961 aaatctctag cagggcccgt ttcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4021 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4081 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4141 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4201 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4261 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4321 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4381 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4441 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4501 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4561 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4621 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4681 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4741 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4801 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4861 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4921 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4981 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5041 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5101 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5161 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5221 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5281 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5341 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5401 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5461 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5521 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5581 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5641 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5701 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5761 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     5821 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     5881 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     5941 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6001 gcttcgcgat gtacgggcca gatatacgcg ctgtggaatg tgtgtcagtt agggtgtgga
     6061 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6301 gccgcctctg cctctgagc
//
