LOCUS       sequence-147832-        4211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for cloning and propagation in DH5alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4211)
  AUTHORS   Li YL, Chandrasekaran V, Carter SD, Woodward CL, Christensen DE, 
            Dryden KA, Pornillos O, Yeager M, Ganser-Pornillos BK, Jensen GJ, 
            Sundquist WI
  TITLE     Primate TRIM5 proteins form hexagonal nets on HIV-1 capsids.
  JOURNAL   Elife. 2016 Jun 2;5. pii: e16269. doi: 10.7554/eLife.16269.
  PUBMED    27253068
REFERENCE   2  (bases 1 to 4211)
  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"; journalName: "Elife."; 
            date: "2016-06-2"; volume: "5. pii: e16269. doi"; pages: " 
            10.7554/eLife.16269"
COMMENT     Sequence Label: pDONR221-OSF-TRIM5-21R(1-300)
FEATURES             Location/Qualifiers
     source          1..4211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             703..726
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             763..786
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             793..816
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             832..855
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    complement(2336..2435)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2452..2471)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2453..2471)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2476..2492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2476..2492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2594..2613)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2605..3414
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2672..2691)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3561..4149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4050..4069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc cgatcgacgg tatcgatgcc accatggcta gctggagcca cccgcagttc
      721 gagaaaggtg gaggtgcccg aggtggatcg ggaggtggat cgtggagcca cccgcagttc
      781 gaaaaaggat tcgattacaa ggatgacgat gacaagggta caagatctcc gctggaagtc
      841 ctgttccagg gccctatggc ttcagcagca cgcttgacaa tgatgtggga ggaggtcaca
      901 tgccctatct gcctggaccc cttcgtggag cctgtgagca tcgagtgtgg ccacagcttc
      961 tgccaggaat gcatctctca ggttgggaaa ggtgggggca gcgtctgtcc tgtgtgccgg
     1021 atcagttacc agcctgagaa catacagcct aatcggcatg tagccaacat agtggagaag
     1081 ctcagggagg tcaagttgag cccagaagag ggacagaagg ttgatcactg tgcacgccat
     1141 ggagagaaac tcctactctt ctgtcaggag gacagcaagg tcatttgctg gctttgtgag
     1201 cggtctcagg agcaccgtgg tcaccacact ttcctcatgg aggaggttgc ccaggagtac
     1261 catgtgaagc tccagacagc tctggagatg ctgaggcaga agcagcagga agctgaaaag
     1321 ttggaagctg acatcagaga agagaaagct tcctggaaga ttcaaataga ctacgacaaa
     1381 accaacgtct cggcagattt tgagcaactg agagagatcc tggactggga ggagagcaat
     1441 gagctgcaga acctggagaa ggaggaagaa gacattctga aaagccttac gaagtctgaa
     1501 acggagatgg tgcagcagac ccagtacatg agagagctca tctcagaact ggagcatcgg
     1561 ttgcaggggt caatgatgga tctactgcag ggtgtggatg gcatcattaa aaggattgag
     1621 aacatgacct tgaagaagcc aaaaactttt cacaaaaatc aaaggagagt gtttcgagct
     1681 cctgatctga aaggaatgct agacatgttt agagagctaa cagatgcccg acgctactgg
     1741 tgagatgtga cactggctac aaacaacatt tcgcatgctg tcattgctga agataagaga
     1801 caagtgagct ctcggaaccc acagataatg tatcaggcac cagggacatt atttacgttt
     1861 ccgtcactca cgaatttcaa ttattgtact ggcgtcctgg gctcccaaag tatcacatca
     1921 gggaagcatt actgggaggt agatgtgtcc aagaaaagtg cttggatcct gggggtatgt
     1981 gctggcttcc aatccgatgc aatgtataat attgaacaaa atgaaaatta tcaacctaaa
     2041 tatggctact gggttatagg gttacaggaa ggagttaaat atagtgtttt ccaggatggt
     2101 tcctcacata ctccttttgc tcctttcatt gtgcccctct ctgtgattat ttgtcctgat
     2161 cgtgttggag ttttcgtaga ctatgaggct tgcactgtct cattcttcaa tatcacaaac
     2221 catggatttc tcatctataa gttttctcag tgttcttttt ctaagcctgt atttccatat
     2281 ttaaatccca gaaaatgtac agtccccatg actctgtgct caccaagctc ttaggaccca
     2341 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     2401 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     2461 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     2521 tgatgttaca ttgcacaaga taaaataata tcatcatgaa caataaaact gtctgcttac
     2581 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     2641 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     2701 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     2761 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     2821 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     2881 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     2941 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     3001 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     3061 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     3121 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     3181 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     3241 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     3301 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     3361 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     3421 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     3481 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     3541 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3601 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3661 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3721 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3781 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3841 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3901 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3961 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4021 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4081 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4141 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4201 gctcacatgt t
//
