LOCUS       sequence-259345-        8011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one huTRIM5 rescue-TRIM5 shRNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8011)
  AUTHORS   Kim K, Dauphin A, Komurlu S, McCauley SM, Yurkovetskiy L, Carbone C,
            Diehl WE, Strambio-De-Castillia C, Campbell EM, Luban J
  TITLE     Cyclophilin A protects HIV-1 from restriction by human TRIM5alpha.
  JOURNAL   Nat Microbiol. 2019 Dec;4(12):2044-2051. doi: 
            10.1038/s41564-019-0592-5. Epub 2019 Oct 21.
  PUBMED    31636416
REFERENCE   2  (bases 1 to 8011)
  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/s41564-019-0592-5"; journalName: "Nat Microbiol"; date: 
            "2019-12"; volume: "4"; issue: "12"; pages: "2044-2051"
COMMENT     Sequence Label: pPU-huTRIMmiR30-TRIM5
FEATURES             Location/Qualifiers
     source          1..8011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..79)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        64..393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     306..325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        405..784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        786..984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     939..959
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1002..1182
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1229..1354
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1851..2084
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2269..2313
                     /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    2611..2728
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2865..3272
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3287..3883
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3287..3306)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3783..3803
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3884..4111
                     /codon_start=1
                     /product="human ubiquitin variant in which Lys-48 has been 
                     mutated to arginine"
                     /label=ubiquitin-K48R
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     ncRNA           5679..5693
                     /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             5827..6007
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6078..6666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6158..6177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6837..7628,7629..7697))
                     /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     7460..7479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7698..7802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7877..7896)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgctgtgg
       61 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
      361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcg cgttgacatt gattattgac
      421 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      481 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      541 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      601 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      661 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      721 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      781 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      841 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      901 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      961 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     1021 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     1081 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     1141 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1201 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1261 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1321 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1381 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1441 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1501 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1561 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1621 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1681 cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1741 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1801 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1861 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1921 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1981 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2041 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2101 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2161 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2221 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2281 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2341 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2401 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2461 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2521 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2581 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2641 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2701 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     2761 ttggttaatt aactgcagcc ccgataaaat aaaagatttt atttagtctc cagaaaaagg
     2821 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc tgcagtaacg ccattttgca
     2881 aggcatggaa aaataccaaa ccaagaatag agaagttcag atcaagggcg ggtacatgaa
     2941 aatagctaac gttgggccaa acaggatatc tgcggtgagc agtttcggcc ccggcccggg
     3001 gccaagaaca gatggtcacc gcagtttcgg ccccggcccg aggccaagaa cagatggtcc
     3061 ccagatatgg cccaaccctc agcagtttct taagacccat cagatgtttc caggctcccc
     3121 caaggacctg aaatgaccct gcgccttatt tgaattaacc aatcagcctg cttctcgctt
     3181 ctgttcgcgc gcttctgctt cccgagctct ataaaagagc tcacaacccc tcactcggcg
     3241 cgccagtcct ccgacagact gagtcgcccg ggggtctagt gccaccatga ccgagtacaa
     3301 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     3361 cgcgttcgcc gactaccccg ccacgcgcca caccgtggac ccggaccgcc acatcgagcg
     3421 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3481 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3541 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3601 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3661 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3721 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3781 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3841 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccatgcaga tctttgtgaa
     3901 gaccctcact ggcaaaacca tcacccttga ggtcgagccc agtgacacca ttgagaatgt
     3961 caaagccaaa attcaagaca aggagggtat cccacctgac cagcagcgtc tgatatttgc
     4021 cggcagacag ctggaggatg gccgcactct ctcagactac aacatccaga aagagtccac
     4081 cctgcacctg gtgttgcgcc tgcgaggtgg catggcttct gggatcctgg ttaatgtaaa
     4141 ggaggaggtg acctgcccca tctgcctgga actcctgaca caacccctga gcctggactg
     4201 cggccacagc ttctgtcagg cctgtttaac agctaaccac aagaagtcca tgctagacaa
     4261 aggagagagt agctgccctg tgtgccggat cagttaccag cctgagaaca tacggcctaa
     4321 tcggcatgta gccaacatag tggagaagct cagggaggtc aagttgagcc cagaggggca
     4381 gaaagttgat cattgtgcac gccatggaga gaaacttcta ctcttctgtc aggaggacgg
     4441 gaaggtcatt tgctggcttt gtgagcggtc tcaggagcac cgtggtcacc acacgttcct
     4501 cacagaggag gttgcccggg agtaccaagt gaagctccag gcagctctgg agatgctgag
     4561 gcagaagcag caggaagctg aagagttgga agctgacatc agagaagaga aagcttcctg
     4621 gaagactcaa atacagtatg acaaaaccaa cgtcttggca gattttgagc aactgagaga
     4681 catcctggac tgggaggaga gcaatgagct gcaaaacctg gagaaggagg aggaagacat
     4741 tctgaaaagc cttacgaact ctgaaactga gatggtgcag cagacccagt ccctgagaga
     4801 gctcatctca gatctggagc atcggctgca ggggtcagtg atggagctgc ttcagggtgt
     4861 ggatggcgtc ataaaaagga cggagaacgt gaccttgaag aagccagaaa cttttccaaa
     4921 aaatcaaagg agagtgtttc gagctcctga tctgaaagga atgctagaag tgtttagaga
     4981 gctgacagat gtccgacgct actgggttga tgtgacagtg gctccaaaca acatttcatg
     5041 tgctgtcatt tctgaagata agagacaagt gagctctccg aaaccacaga taatatatgg
     5101 ggcacgaggg acaagatacc agacatttgt gaatttcaat tattgtactg gcatcctggg
     5161 ctctcaaagt atcacatcag ggaaacatta ctgggaggta gacgtgtcca agaaaactgc
     5221 ttggatactg ggggtatgtg ctggcttcca acctgatgca atgtgtaata ttgaaaaaaa
     5281 tgaaaattat caacctaaat acggctactg ggttataggg ttagaggaag gagttaaatg
     5341 tagtgctttc caggatagtt ccttccatac tccttctgtt cctttcattg tgcccctctc
     5401 tgtgattatt tgtcctgatc gtgttggagt tttcctagac tatgaggctt gcactgtctc
     5461 attcttcaat atcacaaacc atggatttct catctataag ttttctcact gttctttttc
     5521 tcagcctgta tttccatatt taaatcctag aaaatgtgga gtccccatga ctctgtgctc
     5581 accaagctct tgagcggccg ccttcttaac ccaacagaag gctcgagaag gtatattgct
     5641 gttgacagtg agcgcgccaa gcatgcctca ctgcaatagt gaagccacag atgtattgca
     5701 gtgaggcatg cttggcatgc ctactgcctc ggacttcaag gggctagaat tcggcagctg
     5761 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaact gcatccggac
     5821 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5881 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5941 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6001 tctagcaggg cccgtttcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6241 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6481 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6721 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6781 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7801 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7861 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7921 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7981 aaggcttgac cgacaattgc atgaagaatc t
//
