LOCUS       Exported                3809 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAZ1LT7, complete sequence.
ACCESSION   KU513487
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3809)
  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 3809)
  AUTHORS   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 3809)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3809
                     /organism="Cloning vector pAZ1LT7"
                     /mol_type="other DNA"
                     /db_xref="taxon:1873717"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             457..479
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            487..1563
                     /gene="hly-10LT7"
                     /label=hly-10LT7
     CDS             487..1560
                     /codon_start=1
                     /gene="hly-10LT7"
                     /product="interleukin-10/OmpF fusion protein"
                     /label=hly-10LT7
                     /note="human interleukin-10 fusion protein with ompF 
                     leader"
                     /protein_id="ANV82811.1"
                     /translation="MMKRNILAVIVPALLVAGTANASPGQGTQSENSCTHFPGNLPNML
                     RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN
                     QDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS
                     EFDIFINYIEAYMTMKIRNGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRD
                     LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQD
                     PDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEF
                     DIFINYIEAYMTMKIRN"
     primer_bind     complement(1588..1604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1612..1628
                     /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)."
     promoter        complement(1636..1666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1681..1702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1990..2578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2749..3609)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="ANV82812.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2749..3609)
                     /gene="bla"
                     /label=bla
     promoter        complement(3610..3714)
                     /gene="bla"
                     /label=AmpR promoter