LOCUS       sequence-343467-        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-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    487..720
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             905..949
                     /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    1247..1364
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1501..1908
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           2623..2637
                     /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             2771..2951
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3022..3610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3102..3121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3781..4572,4573..4641))
                     /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     4404..4423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4642..4746)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4821..4840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5014..5034)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5019..5348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5199..5334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5261..5280
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5360..5739
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5741..5939
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5894..5914
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5957..6137
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6184..6309
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
       61 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
      121 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
      181 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
      241 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
      301 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
      361 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
      421 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
      481 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
      541 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
      601 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
      661 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
      721 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
      781 ttggagtaat aaatctctgg aacagatttg gaatcacacg acttggatgg agtgggacag
      841 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
      901 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
      961 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     1021 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     1081 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     1141 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     1201 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     1261 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1321 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     1381 acagcagaga tccagtttgg ttaattaact gcagccccga taaaataaaa gattttattt
     1441 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagctgca
     1501 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
     1561 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
     1621 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
     1681 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
     1741 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
     1801 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
     1861 aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccggggg tctagaacgc
     1921 gtgccgccat gaccgaatac aaacctaccg tgaggctggc tacaagagat gatgtcccaa
     1981 gggctgtgag aacactggcc gccgcttttg ccgattaccc tgccacacgc cacactgtgg
     2041 acccagatcg gcatatcgag agagtgactg agctgcagga actgttcctg acccgagtgg
     2101 gcctggacat tgggaaggtc tgggtcgcag acgatggagc agctgtggct gtctggacca
     2161 caccagagag cgtggaagcc ggagctgtct ttgcagagat cggccctaga atggcagaac
     2221 tgagcggctc caggctggca gcacagcagc agatggaggg actgctggcc ccacacaggc
     2281 ctaaggaacc agcatggttc ctggctaccg tgggggtctc tcctgaccat cagggcaaag
     2341 gactgggaag tgctgtggtc ctgccaggag tggaggctgc agaacgagct ggagtccctg
     2401 catttctgga gacctctgct ccacgaaacc tgcccttcta tgaacggctg ggctttactg
     2461 tgaccgcaga tgtggaggtc cccgaaggac ctaggacctg gtgcatgaca cgcaaacccg
     2521 gcgcctgagc gatcgccgcg gccgccttct taacccaaca gaaggctcga gaaggtatat
     2581 tgctgttgac agtgagcgca gtaactattc ccaaagatta tagtgaagcc acagatgtat
     2641 aatctttggg aatagttact ttgcctactg cctcggactt caaggggcta gaattcggca
     2701 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aactgcatcc
     2761 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     2821 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     2881 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     2941 aatctctagc agggcccgtt tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4741 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     4801 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     4861 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     4921 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     4981 cttcgcgatg tacgggccag atatacgcgc tgtggaatgt gtgtcagtta gggtgtggaa
     5041 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5101 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5161 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5221 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5281 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5341 tttgcaaaaa gctcgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     5401 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5461 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5521 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5581 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5641 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5701 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5761 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     5821 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     5881 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
     5941 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6001 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6061 gtgtgcccgt ctgttgtgag actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6121 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     6181 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     6241 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     6301 gagagcgtca gtattaagc
//
