LOCUS       sequence-343446-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HNRNPR 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-HNRNPR-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        33..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             630..810
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    857..982
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1479..1712
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1897..1941
                     /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    2239..2356
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2493..2900
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3615..3629
                     /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             3763..3943
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4014..4602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4094..4113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4773..5564,5565..5633))
                     /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     5396..5415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5634..5738)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5813..5832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6006..6026)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6011..6319,1..21)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6191..6319,1..7)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6253..6272
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ggaggcctag gcttttgcaa aaagctcgcg ttgacattga ttattgacta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      661 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      721 tgcttcaagt agtgtgtgcc cgtctgttgt gagactctgg taactagaga tccctcagac
      781 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      841 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      901 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      961 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1021 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1081 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1141 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1201 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1261 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1321 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1381 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1441 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1501 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1561 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1621 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1681 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1741 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1801 tggaatcaca cgacttggat ggagtgggac agagaaatta acaattacac aagcttaata
     1861 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1921 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1981 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     2041 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     2101 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     2161 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     2221 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2281 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2341 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa
     2401 ctgcagcccc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     2461 accccacctg taggtttggc aagctagctg cagtaacgcc attttgcaag gcatggaaaa
     2521 ataccaaacc aagaatagag aagttcagat caagggcggg tacatgaaaa tagctaacgt
     2581 tgggccaaac aggatatctg cggtgagcag tttcggcccc ggcccggggc caagaacaga
     2641 tggtcaccgc agtttcggcc ccggcccgag gccaagaaca gatggtcccc agatatggcc
     2701 caaccctcag cagtttctta agacccatca gatgtttcca ggctccccca aggacctgaa
     2761 atgaccctgc gccttatttg aattaaccaa tcagcctgct tctcgcttct gttcgcgcgc
     2821 ttctgcttcc cgagctctat aaaagagctc acaacccctc actcggcgcg ccagtcctcc
     2881 gacagactga gtcgcccggg ggtctagaac gcgtgccgcc atgaccgaat acaaacctac
     2941 cgtgaggctg gctacaagag atgatgtccc aagggctgtg agaacactgg ccgccgcttt
     3001 tgccgattac cctgccacac gccacactgt ggacccagat cggcatatcg agagagtgac
     3061 tgagctgcag gaactgttcc tgacccgagt gggcctggac attgggaagg tctgggtcgc
     3121 agacgatgga gcagctgtgg ctgtctggac cacaccagag agcgtggaag ccggagctgt
     3181 ctttgcagag atcggcccta gaatggcaga actgagcggc tccaggctgg cagcacagca
     3241 gcagatggag ggactgctgg ccccacacag gcctaaggaa ccagcatggt tcctggctac
     3301 cgtgggggtc tctcctgacc atcagggcaa aggactggga agtgctgtgg tcctgccagg
     3361 agtggaggct gcagaacgag ctggagtccc tgcatttctg gagacctctg ctccacgaaa
     3421 cctgcccttc tatgaacggc tgggctttac tgtgaccgca gatgtggagg tccccgaagg
     3481 acctaggacc tggtgcatga cacgcaaacc cggcgcctga gcgatcgccg cggccgcctt
     3541 cttaacccaa cagaaggctc gagaaggtat attgctgttg acagtgagcg caactgtgtg
     3601 acagctatga aatagtgaag ccacagatgt atttcatagc tgtcacacag ttttgcctac
     3661 tgcctcggac ttcaaggggc tagaattcgg cagctgtaga tcttagccac tttttaaaag
     3721 aaaagggggg actggaaggg ctaactgcat ccggactgta ctgggtctct ctggttagac
     3781 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     3841 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     3901 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttcatgtga
     3961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5761 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     5821 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     5881 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     5941 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     6001 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6061 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6181 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6241 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     6301 gtagtgagga ggctttttt
//
