LOCUS       sequence-343781-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF2AK2 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-EIF2AK2-ts1
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     546..566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             609..789
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    836..961
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1458..1691
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1876..1920
                     /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    2218..2335
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2472..2879
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3594..3608
                     /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             3742..3922
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3993..4581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4073..4092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4752..5543,5544..5612))
                     /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     5375..5394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5613..5717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5792..5811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5985..6005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5990..6319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6170..6305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6232..6251
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aagctcgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg
      601 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
      661 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
      721 gtctgttgtg agactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
      781 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
      841 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
      901 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
      961 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     1021 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     1081 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     1141 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     1201 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     1261 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga
     1321 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1381 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1441 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1501 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1561 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1621 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1681 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     1741 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacttggatg
     1801 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     1861 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     1921 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     1981 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     2041 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     2101 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     2161 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
     2221 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2281 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2341 ttattacagg gacagcagag atccagtttg gttaattaac tgcagccccg ataaaataaa
     2401 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     2461 agctagctgc agtaacgcca ttttgcaagg catggaaaaa taccaaacca agaatagaga
     2521 agttcagatc aagggcgggt acatgaaaat agctaacgtt gggccaaaca ggatatctgc
     2581 ggtgagcagt ttcggccccg gcccggggcc aagaacagat ggtcaccgca gtttcggccc
     2641 cggcccgagg ccaagaacag atggtcccca gatatggccc aaccctcagc agtttcttaa
     2701 gacccatcag atgtttccag gctcccccaa ggacctgaaa tgaccctgcg ccttatttga
     2761 attaaccaat cagcctgctt ctcgcttctg ttcgcgcgct tctgcttccc gagctctata
     2821 aaagagctca caacccctca ctcggcgcgc cagtcctccg acagactgag tcgcccgggg
     2881 gtctagaacg cgtgccgcca tgaccgaata caaacctacc gtgaggctgg ctacaagaga
     2941 tgatgtccca agggctgtga gaacactggc cgccgctttt gccgattacc ctgccacacg
     3001 ccacactgtg gacccagatc ggcatatcga gagagtgact gagctgcagg aactgttcct
     3061 gacccgagtg ggcctggaca ttgggaaggt ctgggtcgca gacgatggag cagctgtggc
     3121 tgtctggacc acaccagaga gcgtggaagc cggagctgtc tttgcagaga tcggccctag
     3181 aatggcagaa ctgagcggct ccaggctggc agcacagcag cagatggagg gactgctggc
     3241 cccacacagg cctaaggaac cagcatggtt cctggctacc gtgggggtct ctcctgacca
     3301 tcagggcaaa ggactgggaa gtgctgtggt cctgccagga gtggaggctg cagaacgagc
     3361 tggagtccct gcatttctgg agacctctgc tccacgaaac ctgcccttct atgaacggct
     3421 gggctttact gtgaccgcag atgtggaggt ccccgaagga cctaggacct ggtgcatgac
     3481 acgcaaaccc ggcgcctgag cgatcgccgc ggccgccttc ttaacccaac agaaggctcg
     3541 agaaggtata ttgctgttga cagtgagcgc agtagataca aaacaagtaa atagtgaagc
     3601 cacagatgta tttacttgtt ttgtatctac tatgcctact gcctcggact tcaaggggct
     3661 agaattcggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     3721 taactgcatc cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     3781 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     3841 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     3901 tcagtgtgga aaatctctag cagggcccgt ttcatgtgag caaaaggcca gcaaaaggcc
     3961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5701 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     5761 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5821 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     5881 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     5941 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ctgtggaatg tgtgtcagtt
     6001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     6181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6241 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     6301 aggcctaggc ttttgcaaa
//
