LOCUS       40924_17547        3230 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pENTRL2130K-EndoH-TEVH8STREP DNA, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3230)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Biochem. Biotechnol."; date: "2014"; volume: "172"; issue: "8"; 
            pages: "3978-3988"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /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..456
                     /label=attL1
                     /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
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1393..1416
                     /codon_start=1
                     /label=8xHis
                     /note="8xHis affinity tag"
                     /translation="HHHHHHHH"
     CDS             1432..1455
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     protein_bind    complement(1459..1558)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1681..2487
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2580..3168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 gcaggcaact cctaaaaaac cgccaccatg
      481 cggctcacac tattcgcttt cgtactcgct gtgtgtgccc tcgcaagcaa cgctgatgcc
      541 ccggccccgg tgaagcaggg gccgacctcg gtggcctacg tcgaggtgaa caacaacagc
      601 atgctcaacg tgggcaagta cactctggct gacggaggtg gcaacgcctt cgacgtggct
      661 gtgatcttcg ccgctaacat caactacgac actggtacca agactgctta cctccacttc
      721 aacgagaacg tgcaacgtgt gctggacaac gccgtgacac agatccgccc actgcagcaa
      781 cagggaatca aggtgctgct cagcgtcctc ggtaaccacc aaggcgctgg attcgccaac
      841 ttcccgtctc aacaggctgc ctccgctttc gctaagcagt tgtccgacgc tgtcgccaag
      901 tacggactgg acggtgttga tttcgacgat gagtacgctg aatacggcaa caacggaacc
      961 gcccagccaa acgattcctc tttcgtccac ttggttactg ctctgagggc caacatgcca
     1021 gacaagatca tctcactcta caacatcgga ccagctgcct cgcgcttgtc ctacggtggc
     1081 gtggacgtgt ccgataagtt cgactacgct tggaacccct actacggtac ttggcaagtc
     1141 cccggcatcg ctctgcctaa ggcccagctc tcccctgctg ccgtggagat cggaaggaca
     1201 agcagatcaa ccgtcgctga tctggcccgt cgcactgttg acgaaggtta cggcgtgtac
     1261 ctcacataca acttggatgg aggtgacagg actgccgatg tttccgcctt cacaagagaa
     1321 ctgtacggta gcgaagctgt cagaactccc tcgagtgcag gcgaaaacct gtacttccag
     1381 ggaagctcca gccaccacca tcaccatcat catcacggag gcggttccgc atggagccac
     1441 ccgcagttcg aaaagtagac ccagctttct tgtacaaagt tggcattata agaaagcatt
     1501 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     1561 atccagctgc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1621 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     1681 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     1741 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     1801 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     1861 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     1921 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     1981 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2041 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2101 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2161 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2221 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2281 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2341 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2401 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2461 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacattat
     2521 tcagattggg ccccgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2581 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2641 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2701 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2761 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2821 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2881 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2941 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3001 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3061 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3121 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3181 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
