LOCUS       sequence-343460-        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-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(641..1432,1433..1501))
                     /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     1264..1283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1502..1606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1681..1700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1874..1894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1879..2208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2059..2194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2121..2140
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        2220..2599
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2601..2799
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2754..2774
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2817..2997
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3044..3169
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3666..3899
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4084..4128
                     /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    4426..4543
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4680..5087
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           5802..5816
                     /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             5950..6130
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(join(6201..6319,1..470))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6281..6300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
       61 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      121 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      181 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      241 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      301 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      361 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      421 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      481 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      541 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      601 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      661 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      721 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      781 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
      841 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
      901 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
      961 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1021 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1081 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1141 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1201 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1261 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1321 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1381 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1441 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1501 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1561 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1621 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1681 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     1741 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     1801 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     1861 atatacgcgc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     1921 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     1981 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2041 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2101 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2161 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg
     2221 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     2281 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     2341 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2401 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2461 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2521 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2581 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2641 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2701 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2761 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
     2821 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     2881 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgag
     2941 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     3001 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     3061 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     3121 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     3181 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     3241 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     3301 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     3361 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     3421 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     3481 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     3541 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     3601 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     3661 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     3721 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     3781 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     3841 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     3901 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     3961 tggagtaata aatctctgga acagatttgg aatcacacga cttggatgga gtgggacaga
     4021 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     4081 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     4141 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     4201 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     4261 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     4321 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     4381 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     4441 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     4501 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     4561 cagcagagat ccagtttggt taattaactg cagccccgat aaaataaaag attttattta
     4621 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagctgcag
     4681 taacgccatt ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa
     4741 gggcgggtac atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt
     4801 cggccccggc ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc
     4861 aagaacagat ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat
     4921 gtttccaggc tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca
     4981 gcctgcttct cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca
     5041 acccctcact cggcgcgcca gtcctccgac agactgagtc gcccgggggt ctagaacgcg
     5101 tgccgccatg accgaataca aacctaccgt gaggctggct acaagagatg atgtcccaag
     5161 ggctgtgaga acactggccg ccgcttttgc cgattaccct gccacacgcc acactgtgga
     5221 cccagatcgg catatcgaga gagtgactga gctgcaggaa ctgttcctga cccgagtggg
     5281 cctggacatt gggaaggtct gggtcgcaga cgatggagca gctgtggctg tctggaccac
     5341 accagagagc gtggaagccg gagctgtctt tgcagagatc ggccctagaa tggcagaact
     5401 gagcggctcc aggctggcag cacagcagca gatggaggga ctgctggccc cacacaggcc
     5461 taaggaacca gcatggttcc tggctaccgt gggggtctct cctgaccatc agggcaaagg
     5521 actgggaagt gctgtggtcc tgccaggagt ggaggctgca gaacgagctg gagtccctgc
     5581 atttctggag acctctgctc cacgaaacct gcccttctat gaacggctgg gctttactgt
     5641 gaccgcagat gtggaggtcc ccgaaggacc taggacctgg tgcatgacac gcaaacccgg
     5701 cgcctgagcg atcgccgcgg ccgccttctt aacccaacag aaggctcgag aaggtatatt
     5761 gctgttgaca gtgagcgcca ggaggtctat gaatataaat agtgaagcca cagatgtatt
     5821 tatattcata gacctcctga tgcctactgc ctcggacttc aaggggctag aattcggcag
     5881 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta actgcatccg
     5941 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6001 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6061 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6121 atctctagca gggcccgttt catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6301 cctggaagct ccctcgtgc
//
