LOCUS       sequence-259346-        6574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one control rescue-TRIM5 shRNA knockdown vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6574)
  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 6574)
  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-Empty-miR30-TRIM5
FEATURES             Location/Qualifiers
     source          1..6574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..55)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        40..369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      220..355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     282..301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        381..760
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        762..960
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     915..935
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             978..1158
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1205..1330
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1827..2060
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2245..2289
                     /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    2587..2704
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2841..3248
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3263..3859
                     /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(3263..3282)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3759..3779
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3860..4087
                     /codon_start=1
                     /product="human ubiquitin variant in which Lys-48 has been 
                     mutated to arginine"
                     /label=ubiquitin-K48R
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             4094..4120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     ncRNA           4218..4232
                     /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             4366..4546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4617..5205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4697..4716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5376..6167,6168..6236))
                     /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     5999..6018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6237..6341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6416..6435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttcgcgat gtacgggcca gatatacgcg ctgtggaatg tgtgtcagtt agggtgtgga
       61 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
      241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      301 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      361 ttttgcaaaa agctcgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      421 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      481 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      541 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      601 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      661 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      721 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      781 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      841 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      901 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      961 cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     1021 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     1081 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     1141 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     1201 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     1261 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     1321 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     1381 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     1441 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1501 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1561 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1621 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg
     1681 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     1741 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     1801 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     1861 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     1921 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     1981 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     2041 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     2101 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     2161 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     2221 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     2281 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     2341 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     2401 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     2461 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     2521 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat
     2581 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2641 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2701 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaact gcagccccga
     2761 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     2821 ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat accaaaccaa
     2881 gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg ggccaaacag
     2941 gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg gtcaccgcag
     3001 tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca accctcagca
     3061 gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat gaccctgcgc
     3121 cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg
     3181 agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga cagactgagt
     3241 cgcccggggg tctagtgcca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3301 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3361 gcgccacacc gtggacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3421 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3481 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3541 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3601 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3661 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3721 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     3781 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3841 gacccgcaag cccggtgcca tgcagatctt tgtgaagacc ctcactggca aaaccatcac
     3901 ccttgaggtc gagcccagtg acaccattga gaatgtcaaa gccaaaattc aagacaagga
     3961 gggtatccca cctgaccagc agcgtctgat atttgccggc agacagctgg aggatggccg
     4021 cactctctca gactacaaca tccagaaaga gtccaccctg cacctggtgt tgcgcctgcg
     4081 aggtggcatg ggttacccat acgatgttcc agattacgct tctagaggat ccgcggccgc
     4141 cttcttaacc caacagaagg ctcgagaagg tatattgctg ttgacagtga gcgcgccaag
     4201 catgcctcac tgcaatagtg aagccacaga tgtattgcag tgaggcatgc ttggcatgcc
     4261 tactgcctcg gacttcaagg ggctagaatt cggcagctgt agatcttagc cactttttaa
     4321 aagaaaaggg gggactggaa gggctaactg catccggact gtactgggtc tctctggtta
     4381 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     4441 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     4501 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttcatg
     4561 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4621 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4681 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4741 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4801 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4861 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4921 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4981 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5041 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5101 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5161 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5221 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5281 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5341 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5401 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5461 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5521 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5581 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5641 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5701 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5761 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5821 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5881 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5941 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6001 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6061 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6121 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6181 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6241 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6301 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6361 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6421 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6481 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     6541 tgaagaatct gcttagggtt aggcgttttg cgct
//
