LOCUS       sequence-8605-ex        6819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with EF1-a promoter, V5-His C-term, Lac Z.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  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 6819)
  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-LacZ (710-1)
FEATURES             Location/Qualifiers
     source          1..6819
                     /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             1895..4939
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPVPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(1897..1919)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4967..5008
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5018..5035
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(5049..5148)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5260..5279)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5271..6080
                     /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(5338..5357)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6170..6758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6659..6678
                     /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 cagatcgaaa cgatgataga tcccgtcgtt ttacaacgtc gtgactggga
     1921 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1981 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2041 atggcgcttt gcctggtttc cggtaccaga agcggtgccg gaaagctggc tggagtgcga
     2101 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     2161 gcccatctac accaacgtaa cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     2221 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     2281 ccagacgcga attatttttg atggcgttaa ctcggcgttt catctgtggt gcaacgggcg
     2341 ctgggtcggt tacggccagg acagtcgttt gccgtctgaa tttgacctga gcgcattttt
     2401 acgcgccgga gaaaaccgcc tcgcggtgat ggtgctgcgt tggagtgacg gcagttatct
     2461 ggaagatcag gatatgtggc ggatgagcgg cattttccgt gacgtctcgt tgctgcataa
     2521 accgactaca caaatcagcg atttccatgt tgccactcgc tttaatgatg atttcagccg
     2581 cgctgtactg gaggctgaag ttcagatgtg cggcgagttg cgtgactacc tacgggtaac
     2641 agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga
     2701 aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa
     2761 cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg aactgcacac
     2821 cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat
     2881 tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca
     2941 cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatatcct
     3001 gctgatgaag cagaacaact ttaacgccgt gcgctgttcg cattatccga accatccgct
     3061 gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg gatgaagcca atattgaaac
     3121 ccacggcatg gtgccaatga atcgtctgac cgatgatccg cgctggctac cggcgatgag
     3181 cgaacgcgta acgcgaatgg tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc
     3241 gctggggaat gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc
     3301 tgtcgatcct tcccgcccgg tgcagtatga aggcggcgga gccgacacca cggccaccga
     3361 tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa
     3421 atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga tcctttgcga
     3481 atacgcccac gcgatgggta acagtcttgg cggtttcgct aaatactggc aggcgtttcg
     3541 tcagtatccc cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa
     3601 atatgatgaa aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa
     3661 cgatcgccag ttctgtatga acggtctggt ctttgccgac cgcacgccgc atccagcgct
     3721 gacggaagca aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga
     3781 agtgaccagc gaatacctgt tccgtcatag cgataacgag ctcctgcact ggatggtggc
     3841 gctggatggt aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa
     3901 acagttgatt gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac
     3961 agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca tcagcgcctg
     4021 gcagcagtgg cgtctggcgg aaaacctcag tgtgacgctc cccgccgcgt cccacgccat
     4081 cccgcatctg accaccagcg aaatggattt ttgcatcgag ctgggtaata agcgttggca
     4141 atttaaccgc cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct
     4201 gacgccgctg cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga
     4261 agcgacccgc attgacccta acgcctgggt cgaacgctgg aaggcggcgg gccattacca
     4321 ggccgaagca gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac
     4381 gaccgctcac gcgtggcagc atcaggggaa aaccttattt atcagccgga aaacctaccg
     4441 gattgatggt agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc
     4501 gcatccggcg cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg
     4561 gctcggatta gggccgcaag aaaactatcc cgaccgcctt actgccgcct gttttgaccg
     4621 ctgggatctg ccattgtcag acatgtatac cccgtacgtc ttcccgagcg aaaacggtct
     4681 gcgctgcggg acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt
     4741 caacatcagc cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca
     4801 cgcggaagaa ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga
     4861 ctcctggagc ccgtcagtat cggcggaatt ccagctgagc gccggtcgct accattacca
     4921 gttggtctgg tgtcaaaaag cggccgctcg aggtcaccca ttcgaaggta agcctatccc
     4981 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     5041 ttatctagac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     5101 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc
     5161 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     5221 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     5281 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     5341 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     5401 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     5461 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     5521 attttatccg tactcctggt gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     5581 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     5641 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     5701 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     5761 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     5821 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     5881 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     5941 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     6001 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     6061 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacattat tcagattggg
     6121 ccccgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6241 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6301 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6361 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6421 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6481 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6541 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6601 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6661 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6721 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6781 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
