LOCUS       Exported                3343 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRhamhIL-10LT, complete sequence.
ACCESSION   KU513488
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3343)
  AUTHORS   Guggenbichler F, Buttner C, Rudolph W, Zimmermann K, Gunzer F, 
            Pohlmann C.
  TITLE     Design of a covalently linked human interleukin-10 fusion protein 
            and its secretory expression in Escherichia coli
  JOURNAL   Appl. Microbiol. Biotechnol. (2016) In press
  PUBMED    27430741
REFERENCE   2  (bases 1 to 3343)
  AUTHORS   Auerbach C, Guggenbichler F, Buettner C, Rudolph W, Zimmermann K, 
            Gunzer F, Poehlmann C.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JAN-2016) Institute of Medical Microbiology and 
            Hygiene, TU Dresden, Fetscherstrasse 74, Dresden, Saxony 01307, 
            Germany
REFERENCE   3  (bases 1 to 3343)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3343
                     /organism="Cloning vector pRhamhIL-10LT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1873718"
     promoter        1..119
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             183..1274
                     /codon_start=1
                     /gene="hIL-10LH"
                     /product="interleukin-10/OmpF fusion protein"
                     /label=hIL-10LH
                     /note="interleukin-10 fusion protein with ompF leder and 
                     c-terminal polyhistidine tag"
                     /protein_id="ANV82813.1"
                     /translation="MMKRNILAVIVPALLVAGTANASPGQGTQSENSCTHFPGNLPNML
                     RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN
                     QDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS
                     EFDIFINYIEAYMTMKIRNGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRD
                     LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQD
                     PDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEF
                     DIFINYIEAYMTMKIRNHHHHHH"
     gene            183..1274
                     /gene="hIL-10LH"
                     /label=hIL-10LH
     CDS             1254..1271
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1303..1350
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1448..1477
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1499..2086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      2098..2125
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2149..2340)
                     /codon_start=1
                     /gene="rop"
                     /product="repressor of prime"
                     /label=rop
                     /protein_id="ANV82814.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            complement(2149..2340)
                     /gene="rop"
                     /label=rop
     CDS             complement(2341..3156)
                     /codon_start=1
                     /gene="aphA-2"
                     /product="aminoglycoside-modifying protein"
                     /label=aphA-2
                     /note="kanamycine resistance protein"
                     /protein_id="ANV82815.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2341..3156)
                     /gene="aphA-2"
                     /label=aphA-2
     promoter        complement(3157..3247)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      3248..3279
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"