LOCUS       sequence-343475-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HNRNPK 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-HNRNPK-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        501..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             717..897
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    944..1069
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1566..1799
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1984..2028
                     /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    2326..2443
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2580..2987
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3702..3716
                     /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             3850..4030
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4101..4689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4181..4200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4860..5651,5652..5720))
                     /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     5483..5502
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5721..5825)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5900..5919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6093..6113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6098..6319,1..108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6278..6319,1..94)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
       61 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg
      121 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
      721 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      781 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgag
      841 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
      901 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
      961 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     1021 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     1081 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     1141 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     1201 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1261 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1321 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1381 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     1441 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1501 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1561 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1621 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1681 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1741 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1801 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1861 tggagtaata aatctctgga acagatttgg aatcacacga cttggatgga gtgggacaga
     1921 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1981 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2041 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2101 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2161 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2221 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2281 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2341 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2401 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2461 cagcagagat ccagtttggt taattaactg cagccccgat aaaataaaag attttattta
     2521 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagctgcag
     2581 taacgccatt ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa
     2641 gggcgggtac atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt
     2701 cggccccggc ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc
     2761 aagaacagat ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat
     2821 gtttccaggc tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca
     2881 gcctgcttct cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca
     2941 acccctcact cggcgcgcca gtcctccgac agactgagtc gcccgggggt ctagaacgcg
     3001 tgccgccatg accgaataca aacctaccgt gaggctggct acaagagatg atgtcccaag
     3061 ggctgtgaga acactggccg ccgcttttgc cgattaccct gccacacgcc acactgtgga
     3121 cccagatcgg catatcgaga gagtgactga gctgcaggaa ctgttcctga cccgagtggg
     3181 cctggacatt gggaaggtct gggtcgcaga cgatggagca gctgtggctg tctggaccac
     3241 accagagagc gtggaagccg gagctgtctt tgcagagatc ggccctagaa tggcagaact
     3301 gagcggctcc aggctggcag cacagcagca gatggaggga ctgctggccc cacacaggcc
     3361 taaggaacca gcatggttcc tggctaccgt gggggtctct cctgaccatc agggcaaagg
     3421 actgggaagt gctgtggtcc tgccaggagt ggaggctgca gaacgagctg gagtccctgc
     3481 atttctggag acctctgctc cacgaaacct gcccttctat gaacggctgg gctttactgt
     3541 gaccgcagat gtggaggtcc ccgaaggacc taggacctgg tgcatgacac gcaaacccgg
     3601 cgcctgagcg atcgccgcgg ccgccttctt aacccaacag aaggctcgag aaggtatatt
     3661 gctgttgaca gtgagcgcta cgatgaaacc tatgattaat agtgaagcca cagatgtatt
     3721 aatcataggt ttcatcgtaa tgcctactgc ctcggacttc aaggggctag aattcggcag
     3781 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta actgcatccg
     3841 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     3901 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     3961 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     4021 atctctagca gggcccgttt catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4501 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4741 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4801 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4861 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4921 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4981 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5041 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5101 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5161 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5221 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5281 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5341 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5401 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5461 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5521 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5581 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5641 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5701 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5761 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5821 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     5881 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     5941 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6001 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     6061 ttcgcgatgt acgggccaga tatacgcgct gtggaatgtg tgtcagttag ggtgtggaaa
     6121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6241 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6301 ttccgcccat tctccgccc
//
