LOCUS       Exported                3230 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pENTRL2130K-EndoH-TEVH8STREP DNA, complete sequence.
ACCESSION   AB838596
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3230)
  AUTHORS   Mitsudome T, Xu J, Nagata Y, Masuda A, Iiyama K, Morokuma D, Li Z, 
            Mon H, Lee JM, Kusakabe T.
  TITLE     Expression, purification, and characterization of 
            endo-beta-N-acetylglucosaminidase H using baculovirus-mediated 
            silkworm protein expression system
  JOURNAL   Appl. Biochem. Biotechnol. 172 (8), 3978-3988 (2014)
  PUBMED    24599668
REFERENCE   2  (bases 1 to 3230)
  AUTHORS   Kusakabe T, Mitsudome T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JUL-2013) Contact:Takahiro Kusakabe Kyushu University,
            Graduate School of Bioresource and Bioenvironmental Sciences, 
            Laboratory of Silkworm Science; Hakozaki 6-10-1, Fukuoka, Fukuoka 
            812-8581, Japan
REFERENCE   3  (bases 1 to 3230)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3230
                     /organism="Cloning vector pENTRL2130K-EndoH-TEVH8STREP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1367419"
     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"
     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"
     misc_feature    457..477
                     /label=L21 enhancer sequence
                     /note="L21 enhancer sequence"
     misc_feature    478..537
                     /label=Bombyx mori 30K protein signal peptide
                     /note="Bombyx mori 30K protein signal peptide"
     misc_feature    538..1362
                     /gene="Endo-H-TEVH8STREP"
                     /label=endo-beta-N-acetylglucosaminidase H-TEVH8STREP
                     /note="endo-beta-N-acetylglucosaminidase H-TEVH8STREP"
     gene            538..1362
                     /gene="Endo-H-TEVH8STREP"
                     /label=Endo-H-TEVH8STREP
     CDS             610..1362
                     /codon_start=1
                     /gene="Endo-H"
                     /product="endo-beta-N-acetylglucosaminidase H"
                     /label=Endo-H
                     /protein_id="BAN66718.1"
                     /translation="VGKYTLADGGGNAFDVAVIFAANINYDTGTKTAYLHFNENVQRVL
                     DNAVTQIRPLQQQGIKVLLSVLGNHQGAGFANFPSQQAASAFAKQLSDAVAKYGLDGVD
                     FDDEYAEYGNNGTAQPNDSSFVHLVTALRANMPDKIISLYNIGPAASRLSYGGVDVSDK
                     FDYAWNPYYGTWQVPGIALPKAQLSPAAVEIGRTSRSTVADLARRTVDEGYGVYLTYNL
                     DGGDRTADVSAFTRELYGSEAVRTPSSAG"
     gene            610..1362
                     /gene="Endo-H"
                     /label=Endo-H
     CDS             1363..1383
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    1393..1416
                     /label=Hisx8 tag sequence
                     /note="Hisx8 tag sequence"
     CDS             1393..1416
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     misc_feature    1432..1458
                     /label=Strep tag sequence
                     /note="Strep tag sequence"
     CDS             1432..1455
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(1459..1558)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1681..2490
                     /codon_start=1
                     /gene="Kan"
                     /product="kanamycin resistance protein"
                     /label=Kan
                     /protein_id="BAN64147.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            1681..2490
                     /gene="Kan"
                     /label=Kan
     rep_origin      2580..3168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"