LOCUS       sequence-251150-        7189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAPM knockdown vector with Myd88 stuffer sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7189)
  AUTHORS   Yurkovetskiy L, Guney MH, Kim K, Goh SL, McCauley S, Dauphin A, 
            Diehl WE, Luban J
  TITLE     Primate immunodeficiency virus proteins Vpx and Vpr counteract 
            transcriptional repression of proviruses by the HUSH complex.
  JOURNAL   Nat Microbiol. 2018 Dec;3(12):1354-1361. doi: 
            10.1038/s41564-018-0256-x. Epub 2018 Oct 8.
  PUBMED    30297740
REFERENCE   2  (bases 1 to 7189)
  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"; doi: 
            "10.1038/s41564-018-0256-x"; journalName: "Nat Microbiol"; date: 
            "2018-12"; volume: "3"; issue: "12"; pages: "1354-1361"
COMMENT     Sequence Label: pAPM D4 BsmBI Myd88 stuffer
FEATURES             Location/Qualifiers
     source          1..7189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(141..161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        146..475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      326..461
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     388..407
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        487..866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        868..1066
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1021..1041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1084..1264
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1311..1436
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1933..2166
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2351..2395
                     /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    2693..2810
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2947..3354
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      4069..4078
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5087..5267
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5338..5926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5418..5437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6097..6888,6889..6957))
                     /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     6720..6739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6958..7062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7137..7156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
       61 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      121 gggccagata tacgcgctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
      421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      481 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      541 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      601 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      661 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      721 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      781 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      841 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      901 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      961 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1021 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
     1081 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     1141 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     1201 ttgtgagact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1261 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1321 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1381 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1441 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1501 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1561 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1621 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1681 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1741 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
     1801 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     1861 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     1921 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     1981 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     2041 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     2101 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     2161 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     2221 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgactt ggatggagtg
     2281 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     2341 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2401 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2461 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2521 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     2581 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     2641 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     2701 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2761 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2821 acagggacag cagagatcca gtttggttaa ttaactgcag ccccgataaa ataaaagatt
     2881 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     2941 gctgcagtaa cgccattttg caaggcatgg aaaaatacca aaccaagaat agagaagttc
     3001 agatcaaggg cgggtacatg aaaatagcta acgttgggcc aaacaggata tctgcggtga
     3061 gcagtttcgg ccccggcccg gggccaagaa cagatggtca ccgcagtttc ggccccggcc
     3121 cgaggccaag aacagatggt ccccagatat ggcccaaccc tcagcagttt cttaagaccc
     3181 atcagatgtt tccaggctcc cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa
     3241 ccaatcagcc tgcttctcgc ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga
     3301 gctcacaacc cctcactcgg cgcgccagtc ctccgacaga ctgagtcgcc cgggggtcta
     3361 gaacgcgtgc cgccatgacc gaatacaaac ctaccgtgag gctggctaca agagatgatg
     3421 tcccaagggc tgtgagaaca ctggccgccg cttttgccga ttaccctgcc acacgccaca
     3481 ctgtggaccc agatcggcat atcgagagag tgactgagct gcaggaactg ttcctgaccc
     3541 gagtgggcct ggacattggg aaggtctggg tcgcagacga tggagcagct gtggctgtct
     3601 ggaccacacc agagagcgtg gaagccggag ctgtctttgc agagatcggc cctagaatgg
     3661 cagaactgag cggctccagg ctggcagcac agcagcagat ggagggactg ctggccccac
     3721 acaggcctaa ggaaccagca tggttcctgg ctaccgtggg ggtctctcct gaccatcagg
     3781 gcaaaggact gggaagtgct gtggtcctgc caggagtgga ggctgcagaa cgagctggag
     3841 tccctgcatt tctggagacc tctgctccac gaaacctgcc cttctatgaa cggctgggct
     3901 ttactgtgac cgcagatgtg gaggtccccg aaggacctag gacctggtgc atgacacgca
     3961 aacccggcgc ctgagcgatc gccgcggccg ccttcttaac ccaacagaag gctcgagaag
     4021 gtatattgct gttgacagtg agcggagacg ggccgcgtct tcggatccgc cgccatggca
     4081 gctggcggac caggcgcagg tagtgctgcg ccagtttctt caacttcctc cctgcccctg
     4141 gctgcgctta acatgcgcgt gagaaggagg ctctccctct tcctgaacgt aagaactcag
     4201 gtggccgcag attggactgc actggctgag gagatggact tcgaatatct ggagatacgg
     4261 cagcttgaaa cccaagctga ccctacagga cggctgctcg acgcctggca gggccgcccc
     4321 ggtgcctctg taggcaggct tctggagctg ctgacgaagc tggggaggga tgatgtgctg
     4381 ctggagttgg gacccagcat tgaggaggat tgtcagaagt atattctgaa gcaacagcag
     4441 gaggaggcgg agaagccact gcaggtcgcg gcagtcgatt ctagtgttcc acggactgcg
     4501 gaacttgcgg gtattaccac actcgatgat cccttgggac acatgccgga gcgctttgac
     4561 gccttcatat gctattgccc cagtgatata cagtttgtgc aggaaatgat tagacaactg
     4621 gagcaaacaa attatcgact gaagctgtgc gtgtccgata gggacgttct tccaggtacc
     4681 tgtgtttgga gtatcgcttc agagctcatc gagaagcggt gtagaagaat ggtggttgta
     4741 gtgagcgacg attatctcca gtcaaaggag tgtgatttcc aaaccaaatt cgcactttcc
     4801 ctcagcccag gggcacatca aaagagactg ataccgataa aatataaagc tatgaagaaa
     4861 gagtttccgt ccattttgag atttatcact gtttgcgact acactaaccc ttgcacgaag
     4921 tcttggtttt ggacacgcct ggctaaagct ctgtcattgc cgtgatggcc acgtctctgc
     4981 ctactgcctc ggacttcaag gggctagaat tcggcagctg tagatcttag ccacttttta
     5041 aaagaaaagg ggggactgga agggctaact gcatccggac tgtactgggt ctctctggtt
     5101 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5161 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5221 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttcat
     5281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6241 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7081 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     7141 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtg
//
