LOCUS       sequence-8632-ex        2972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  modified pENTR vector for C-terminal GST tag fusion with your gene 
            of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2972)
  AUTHORS   Dubin MJ, Bowler C, Benvenuto G
  TITLE     A modified Gateway cloning strategy for overexpressing tagged 
            proteins in plants.
  JOURNAL   Plant Methods. 2008 Jan 22. 4(1):3.
  PUBMED    18211686
REFERENCE   2  (bases 1 to 2972)
  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: "Plant 
            Methods. 2008 Jan 22. 4(1):3."
COMMENT     Sequence Label: pE4c
FEATURES             Location/Qualifiers
     source          1..2972
                     /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"
     CDS             524..1177
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1135..1157
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    complement(1201..1300)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1412..1431)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1423..2232
                     /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
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1490..1509)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2322..2910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2811..2830
                     /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 gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt acagatttcg gacatgcggc cgcaagcgga ggcatgtccc ctatactagg
      541 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
      601 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
      661 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
      721 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
      781 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
      841 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
      901 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
      961 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
     1021 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
     1081 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
     1141 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gattagctcg agatatctag
     1201 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1261 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gcagctctgg
     1321 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1381 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1441 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1501 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     1561 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1621 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1681 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     1741 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgtccctg
     1801 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1861 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1921 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     1981 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2041 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2101 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2161 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2221 gagtttttct aatcagaatt ggttaattgg ttgtaacatt attcagattg ggccccgttc
     2281 cactgagcgt cagacccggt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2341 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2401 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2461 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2521 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2581 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2641 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2701 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2761 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2821 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2881 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2941 ctggcctttt gctggccttt tgctcacatg tt
//
