LOCUS       sequence-259347-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivector for TRIM5 knockdown and puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6319)
  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 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"; doi: 
            "10.1038/s41564-019-0592-5"; journalName: "Nat Microbiol"; date: 
            "2019-12"; volume: "4"; issue: "12"; pages: "2044-2051"
COMMENT     Sequence Label: pAPM-D4-miR30-TRIM5
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..27)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(201..221)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        206..535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      386..521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     448..467
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        547..926
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        928..1126
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1081..1101
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1144..1324
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1371..1496
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1993..2226
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2411..2455
                     /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    2753..2870
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3007..3414
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           4129..4143
                     /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             4277..4457
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4608..4627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5287..6078,6079..6147))
                     /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     5910..5929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6148..6252)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
       61 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      121 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      181 gggccagata tacgcgctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      301 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      361 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      421 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
      481 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      541 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      601 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      661 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      721 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      781 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      841 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      901 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      961 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     1021 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1081 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
     1141 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     1201 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     1261 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1321 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1381 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1441 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1501 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1561 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1621 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1681 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1741 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1801 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
     1861 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     1921 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     1981 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     2041 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     2101 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     2161 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     2221 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     2281 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     2341 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     2401 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2461 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2521 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2581 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     2641 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     2701 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     2761 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2821 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2881 acagggacag cagagatcca gtttggttaa ttaactgcag ccccgataaa ataaaagatt
     2941 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     3001 gctgcagtaa cgccattttg caaggcatgg aaaaatacca aaccaagaat agagaagttc
     3061 agatcaaggg cgggtacatg aaaatagcta acgttgggcc aaacaggata tctgcggtga
     3121 gcagtttcgg ccccggcccg gggccaagaa cagatggtca ccgcagtttc ggccccggcc
     3181 cgaggccaag aacagatggt ccccagatat ggcccaaccc tcagcagttt cttaagaccc
     3241 atcagatgtt tccaggctcc cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa
     3301 ccaatcagcc tgcttctcgc ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga
     3361 gctcacaacc cctcactcgg cgcgccagtc ctccgacaga ctgagtcgcc cgggggtcta
     3421 gaacgcgtgc cgccatgacc gaatacaaac ctaccgtgag gctggctaca agagatgatg
     3481 tcccaagggc tgtgagaaca ctggccgccg cttttgccga ttaccctgcc acacgccaca
     3541 ctgtggaccc agatcggcat atcgagagag tgactgagct gcaggaactg ttcctgaccc
     3601 gagtgggcct ggacattggg aaggtctggg tcgcagacga tggagcagct gtggctgtct
     3661 ggaccacacc agagagcgtg gaagccggag ctgtctttgc agagatcggc cctagaatgg
     3721 cagaactgag cggctccagg ctggcagcac agcagcagat ggagggactg ctggccccac
     3781 acaggcctaa ggaaccagca tggttcctgg ctaccgtggg ggtctctcct gaccatcagg
     3841 gcaaaggact gggaagtgct gtggtcctgc caggagtgga ggctgcagaa cgagctggag
     3901 tccctgcatt tctggagacc tctgctccac gaaacctgcc cttctatgaa cggctgggct
     3961 ttactgtgac cgcagatgtg gaggtccccg aaggacctag gacctggtgc atgacacgca
     4021 aacccggcgc ctgagcgatc gccgcggccg ccttcttaac ccaacagaag gctcgagaag
     4081 gtatattgct gttgacagtg agcgcgccaa gcatgcctca ctgcaatagt gaagccacag
     4141 atgtattgca gtgaggcatg cttggcatgc ctactgcctc ggacttcaag gggctagaat
     4201 tcggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaact
     4261 gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4321 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4381 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4441 gtggaaaatc tctagcaggg cccgtttcat gtgagcaaaa ggccagcaaa aggccaggaa
     4501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6301 cccgatcccc tatggtgca
//
