LOCUS       sequence-343464-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZC3HAV1 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-ZC3HAV1-ts1
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     585..605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             648..828
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    875..1000
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1497..1730
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1915..1959
                     /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    2257..2374
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2511..2918
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3633..3647
                     /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             3781..3961
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4032..4620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4112..4131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4791..5582,5583..5651))
                     /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     5414..5433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5652..5756)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5831..5850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6024..6044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6029..6319,1..39)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6209..6319,1..25)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6271..6290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcgcgtt gacattgatt
       61 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt
      661 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      721 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      781 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      841 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      901 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      961 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     1021 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     1081 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     1141 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     1201 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     1261 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     1321 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg
     1381 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1441 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1501 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1561 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1621 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1681 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1741 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1801 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1861 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1921 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1981 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2041 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2101 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     2161 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     2221 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     2281 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     2341 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     2401 tccagtttgg ttaattaact gcagccccga taaaataaaa gattttattt agtctccaga
     2461 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagctgca gtaacgccat
     2521 tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca agggcgggta
     2581 catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt tcggccccgg
     2641 cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc caagaacaga
     2701 tggtccccag atatggccca accctcagca gtttcttaag acccatcaga tgtttccagg
     2761 ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc
     2821 tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac aacccctcac
     2881 tcggcgcgcc agtcctccga cagactgagt cgcccggggg tctagaacgc gtgccgccat
     2941 gaccgaatac aaacctaccg tgaggctggc tacaagagat gatgtcccaa gggctgtgag
     3001 aacactggcc gccgcttttg ccgattaccc tgccacacgc cacactgtgg acccagatcg
     3061 gcatatcgag agagtgactg agctgcagga actgttcctg acccgagtgg gcctggacat
     3121 tgggaaggtc tgggtcgcag acgatggagc agctgtggct gtctggacca caccagagag
     3181 cgtggaagcc ggagctgtct ttgcagagat cggccctaga atggcagaac tgagcggctc
     3241 caggctggca gcacagcagc agatggaggg actgctggcc ccacacaggc ctaaggaacc
     3301 agcatggttc ctggctaccg tgggggtctc tcctgaccat cagggcaaag gactgggaag
     3361 tgctgtggtc ctgccaggag tggaggctgc agaacgagct ggagtccctg catttctgga
     3421 gacctctgct ccacgaaacc tgcccttcta tgaacggctg ggctttactg tgaccgcaga
     3481 tgtggaggtc cccgaaggac ctaggacctg gtgcatgaca cgcaaacccg gcgcctgagc
     3541 gatcgccgcg gccgccttct taacccaaca gaaggctcga gaaggtatat tgctgttgac
     3601 agtgagcgac ccgagatatg caaaagttat tagtgaagcc acagatgtaa taacttttgc
     3661 atatctcggg ctgcctactg cctcggactt caaggggcta gaattcggca gctgtagatc
     3721 ttagccactt tttaaaagaa aaggggggac tggaagggct aactgcatcc ggactgtact
     3781 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     3841 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     3901 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     3961 agggcccgtt tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4201 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4441 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4741 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4801 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4861 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4921 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4981 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5041 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5101 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5161 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5221 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5281 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5341 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5401 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5461 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5521 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5581 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5641 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5761 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     5821 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     5881 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     5941 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     6001 tacgggccag atatacgcgc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6061 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     6121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6181 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     6301 ctctgagcta ttccagaag
//
