LOCUS       Exported                4366 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pJJDuet30, complete sequence.
ACCESSION   DQ016035
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4366)
  AUTHORS   Zuger S, Iwai H.
  TITLE     Intein-based biosynthetic incorporation of unlabeled protein tags 
            into isotopically labeled proteins for NMR studies
  JOURNAL   Nat. Biotechnol. 23 (6), 736-740 (2005)
  PUBMED    15908942
REFERENCE   2  (bases 1 to 4366)
  AUTHORS   Zueger S, Iwai H.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2005) Chemistry, University of Saskatchewan, 110 
            Science Place, Saskatoon, SK S7N 5C9, Canada
REFERENCE   3  (bases 1 to 4366)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4366
                     /organism="Expression vector pJJDuet30"
                     /mol_type="other DNA"
                     /label=in vivo protein splicing
                     /note="in vivo protein splicing"
                     /db_xref="taxon:328606"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             71..676
                     /codon_start=1
                     /product="protein G/SspDnaE fusion protein"
                     /label=protein G/SspDnaE fusion protein
                     /note="N-terminal domain of split DnaE intein derived from 
                     Synechocystis sp. PCC6803"
                     /protein_id="AAY41168.1"
                     /translation="MGSSHHHHHHSSGLVPRGSHMQYKLILNGKTLKGETTTEAVDAAT
                     AEKVFKQYANDNGVDGEWTYDDATKTYTVTEGSCLSFGTEILTVEYGPLPIGKIVSEEI
                     NCSVYSVDPEGRVYTQAIAQWHDRGEQEVLEYELEDGSVIRATSDHRFLTTDYQLLAIE
                     EIFARQLDLLTLENIKQTEEALDNHRLPFPLLDAGTIK"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        751..769
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    770..794
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             903..947
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      999..1046
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1276..2091)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2092..2183)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2199..2948)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3144..4226)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4227..4304)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4350..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"