LOCUS       sequence-343176-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LRPPRC 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-LRPPRC-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     80..99
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        179..558
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        560..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     713..733
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             776..956
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1003..1128
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1625..1858
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2043..2087
                     /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    2385..2502
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2639..3046
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3761..3775
                     /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             3909..4089
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4160..4748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4240..4259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4919..5710,5711..5779))
                     /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     5542..5561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5780..5884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5959..5978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6152..6172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6157..6319,1..167)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
       61 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
      121 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcgcgttga
      181 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      241 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      301 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      361 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      421 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      481 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      541 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      601 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      661 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      721 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
      781 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      841 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgaga
      901 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      961 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     1021 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1081 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1141 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1201 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     1261 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1321 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1381 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1441 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     1501 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1561 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1621 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1681 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1741 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1801 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1861 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1921 ggagtaataa atctctggaa cagatttgga atcacacgac ttggatggag tgggacagag
     1981 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2041 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2101 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2161 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2221 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2281 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2341 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2401 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2461 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2521 agcagagatc cagtttggtt aattaactgc agccccgata aaataaaaga ttttatttag
     2581 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagctgcagt
     2641 aacgccattt tgcaaggcat ggaaaaatac caaaccaaga atagagaagt tcagatcaag
     2701 ggcgggtaca tgaaaatagc taacgttggg ccaaacagga tatctgcggt gagcagtttc
     2761 ggccccggcc cggggccaag aacagatggt caccgcagtt tcggccccgg cccgaggcca
     2821 agaacagatg gtccccagat atggcccaac cctcagcagt ttcttaagac ccatcagatg
     2881 tttccaggct cccccaagga cctgaaatga ccctgcgcct tatttgaatt aaccaatcag
     2941 cctgcttctc gcttctgttc gcgcgcttct gcttcccgag ctctataaaa gagctcacaa
     3001 cccctcactc ggcgcgccag tcctccgaca gactgagtcg cccgggggtc tagaacgcgt
     3061 gccgccatga ccgaatacaa acctaccgtg aggctggcta caagagatga tgtcccaagg
     3121 gctgtgagaa cactggccgc cgcttttgcc gattaccctg ccacacgcca cactgtggac
     3181 ccagatcggc atatcgagag agtgactgag ctgcaggaac tgttcctgac ccgagtgggc
     3241 ctggacattg ggaaggtctg ggtcgcagac gatggagcag ctgtggctgt ctggaccaca
     3301 ccagagagcg tggaagccgg agctgtcttt gcagagatcg gccctagaat ggcagaactg
     3361 agcggctcca ggctggcagc acagcagcag atggagggac tgctggcccc acacaggcct
     3421 aaggaaccag catggttcct ggctaccgtg ggggtctctc ctgaccatca gggcaaagga
     3481 ctgggaagtg ctgtggtcct gccaggagtg gaggctgcag aacgagctgg agtccctgca
     3541 tttctggaga cctctgctcc acgaaacctg cccttctatg aacggctggg ctttactgtg
     3601 accgcagatg tggaggtccc cgaaggacct aggacctggt gcatgacacg caaacccggc
     3661 gcctgagcga tcgccgcggc cgccttctta acccaacaga aggctcgaga aggtatattg
     3721 ctgttgacag tgagcgatgc cagcaagatt cttggatata gtgaagccac agatgtatat
     3781 ccaagaatct tgctggcact gcctactgcc tcggacttca aggggctaga attcggcagc
     3841 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ctgcatccgg
     3901 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     3961 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     4021 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     4081 tctctagcag ggcccgtttc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4561 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4921 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4981 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5041 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5101 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5161 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5221 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5281 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5341 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5401 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5461 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5521 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5581 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5641 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5701 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5761 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5821 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5881 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     5941 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     6001 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     6061 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     6121 tcgcgatgta cgggccagat atacgcgctg tggaatgtgt gtcagttagg gtgtggaaag
     6181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6301 tagtcagcaa ccatagtcc
//
