LOCUS       sequence-8604-ex        3768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with EF1-a promoter, V5-His C-term, frame C .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3768)
  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 3768)
  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 C (709-1)
FEATURES             Location/Qualifiers
     source          1..3768
                     /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             1916..1957
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1967..1984
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1998..2097)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2209..2228)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2220..3029
                     /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(2287..2306)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3119..3707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3608..3627
                     /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 ggtcacccat tcgaaggtaa
     1921 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     1981 ccattgagtt tatctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2041 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2101 tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     2161 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     2221 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     2281 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     2341 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2401 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2461 ccatcaagca ttttatccgt actcctggtg atgcatggtt actcaccact gcgatccccg
     2521 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2581 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2641 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2701 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2761 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2821 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2881 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2941 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3001 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacattatt
     3061 cagattgggc cccgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3121 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3181 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3241 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3301 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3361 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3421 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3481 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3541 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3601 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3661 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3721 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
