LOCUS       sequence-222777-        3773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with EF1-a promoter, V5-His C-term, frame A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3773)
  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 3773)
  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 A (696-6)
FEATURES             Location/Qualifiers
     source          1..3773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      122..165
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      297..324
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     332..355
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    374..473
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        604..1782
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          835..1773
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1730..1750
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1799..1818
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1799..1817
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1936..1977
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1987..2004
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2018..2117)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2229..2248)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2240..3049
                     /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(2307..2326)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3139..3727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3628..3647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
       61 ctagcatgga tctcggggac gtctaactac taagcgagag tagggaactg ccaggcatca
      121 aataaaacga aaggcccagt cttccgactg agcctttcgt tttatctgtt gtttgtcggt
      181 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg tgaagcaacg
      241 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa
      301 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctgtta gttagttact
      361 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
      421 aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagcag gctccaccat
      481 gggaaccgac attgattatt gactaggctt ttgcaaaaag ctttgcaaag atggataaag
      541 ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
      601 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
      661 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
      721 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
      781 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
      841 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
      901 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      961 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
     1021 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
     1081 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
     1141 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
     1201 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
     1261 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
     1321 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
     1381 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     1441 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     1501 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     1561 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1621 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1681 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1741 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattag cttggtacta
     1801 atacgactca ctatagggag acccaagctg gctaggtaag cttggtaccg agctcggatc
     1861 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct
     1921 agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     1981 accggtcatc atcaccatca ccattgagtt tatctagacc cagctttctt gtacaaagtt
     2041 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2101 aataaaatca ttatttgcca tccagctgca gctctggccc gtgtctcaaa atctctgatg
     2161 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2221 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2281 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2341 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2401 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2461 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctggtg atgcatggtt
     2521 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2581 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2641 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2701 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2761 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2821 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2881 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2941 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3001 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3061 taattggttg taacattatt cagattgggc cccgttccac tgagcgtcag accccgtaga
     3121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggc
//
