LOCUS       sequence-343782-        6319 bp    DNA     circular SYN 15-APR-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 Apr 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-ts3
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     243..262
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        342..721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..921
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     876..896
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             939..1119
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1166..1291
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1788..2021
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2206..2250
                     /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    2548..2665
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2802..3209
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3924..3938
                     /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             4072..4252
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4323..4911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4403..4422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5082..5873,5874..5942))
                     /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     5705..5724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5943..6047)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6122..6141)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(join(6315..6319,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcgcgt tgacattgat tattgactag
      361 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      421 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      481 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      541 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      601 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      661 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      721 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      781 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      841 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      901 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
      961 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1021 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg agactctggt aactagagat
     1081 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     1141 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     1201 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     1261 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     1321 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     1381 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1441 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1501 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1561 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1621 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1681 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1741 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     1801 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     1861 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     1921 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     1981 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     2041 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     2101 gaacagattt ggaatcacac gacttggatg gagtgggaca gagaaattaa caattacaca
     2161 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     2221 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     2281 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     2341 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     2401 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     2461 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
     2521 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     2581 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     2641 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     2701 gttaattaac tgcagccccg ataaaataaa agattttatt tagtctccag aaaaaggggg
     2761 gaatgaaaga ccccacctgt aggtttggca agctagctgc agtaacgcca ttttgcaagg
     2821 catggaaaaa taccaaacca agaatagaga agttcagatc aagggcgggt acatgaaaat
     2881 agctaacgtt gggccaaaca ggatatctgc ggtgagcagt ttcggccccg gcccggggcc
     2941 aagaacagat ggtcaccgca gtttcggccc cggcccgagg ccaagaacag atggtcccca
     3001 gatatggccc aaccctcagc agtttcttaa gacccatcag atgtttccag gctcccccaa
     3061 ggacctgaaa tgaccctgcg ccttatttga attaaccaat cagcctgctt ctcgcttctg
     3121 ttcgcgcgct tctgcttccc gagctctata aaagagctca caacccctca ctcggcgcgc
     3181 cagtcctccg acagactgag tcgcccgggg gtctagaacg cgtgccgcca tgaccgaata
     3241 caaacctacc gtgaggctgg ctacaagaga tgatgtccca agggctgtga gaacactggc
     3301 cgccgctttt gccgattacc ctgccacacg ccacactgtg gacccagatc ggcatatcga
     3361 gagagtgact gagctgcagg aactgttcct gacccgagtg ggcctggaca ttgggaaggt
     3421 ctgggtcgca gacgatggag cagctgtggc tgtctggacc acaccagaga gcgtggaagc
     3481 cggagctgtc tttgcagaga tcggccctag aatggcagaa ctgagcggct ccaggctggc
     3541 agcacagcag cagatggagg gactgctggc cccacacagg cctaaggaac cagcatggtt
     3601 cctggctacc gtgggggtct ctcctgacca tcagggcaaa ggactgggaa gtgctgtggt
     3661 cctgccagga gtggaggctg cagaacgagc tggagtccct gcatttctgg agacctctgc
     3721 tccacgaaac ctgcccttct atgaacggct gggctttact gtgaccgcag atgtggaggt
     3781 ccccgaagga cctaggacct ggtgcatgac acgcaaaccc ggcgcctgag cgatcgccgc
     3841 ggccgccttc ttaacccaac agaaggctcg agaaggtata ttgctgttga cagtgagcgc
     3901 cctgagaaca gcaaaaatag atagtgaagc cacagatgta tctatttttg ctgttctcag
     3961 gatgcctact gcctcggact tcaaggggct agaattcggc agctgtagat cttagccact
     4021 ttttaaaaga aaagggggga ctggaagggc taactgcatc cggactgtac tgggtctctc
     4081 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4141 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4201 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
     4261 ttcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4321 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4381 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4441 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4501 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4561 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4621 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4681 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4741 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4801 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4861 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4921 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4981 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5041 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5101 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5161 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5221 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5281 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5341 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5401 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5461 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5521 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5581 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5641 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5701 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5761 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5821 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5881 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5941 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6001 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6061 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     6121 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     6181 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     6241 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     6301 gatatacgcg ctgtggaat
//
