LOCUS       sequence-8603-ex        3776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with EF1-a promoter, V5-His C-term, frame B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3776)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 3776)
  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: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pEF-ENTR B (708-1)
FEATURES             Location/Qualifiers
     source          1..3776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        588..1766
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          819..1757
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1714..1734
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1783..1802
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1783..1801
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1924..1965
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1975..1992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2006..2105)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2217..2236)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2228..3037
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2295..2314)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3127..3715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3616..3635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac cgacattgat
      481 tattgactag gcttttgcaa aaagctttgc aaagatggat aaagttttaa acagagagga
      541 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
      601 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      661 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      721 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      781 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
      841 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
      901 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      961 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     1021 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     1081 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     1141 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     1201 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     1261 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     1321 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     1381 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     1441 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1501 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1561 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1621 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1681 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1741 tttttcttcc atttcaggtg tcgtgaggaa ttagcttggt actaatacga ctcactatag
     1801 ggagacccaa gctggctagg taagcttggt accgagctcg gatccactag tccagtgtgg
     1861 tggaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
     1921 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat
     1981 caccatcacc attgagttta tctagaccca gctttcttgt acaaagttgg cattataaga
     2041 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     2101 atttgccatc cagctgcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2161 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2221 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2281 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2341 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2401 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2461 tcttccgacc atcaagcatt ttatccgtac tcctggtgat gcatggttac tcaccactgc
     2521 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2581 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2641 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2701 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2761 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2821 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2881 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2941 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3001 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3061 acattattca gattgggccc cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3241 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3481 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
