LOCUS       Exported                9305 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pLEXSY-LC1, complete sequence.
ACCESSION   JF327852
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9305)
  AUTHORS   Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.
  TITLE     High-Throughput Protein Expression Using a Combination of 
            Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein 
            (IFP) Detection
  JOURNAL   PLoS ONE 6 (4), E18900 (2011)
  PUBMED    21541323
REFERENCE   2  (bases 1 to 9305)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 9305)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9305
                     /organism="Expression vector LIC-pLEXSY-LC1"
                     /mol_type="other DNA"
                     /label=Leishmania tarentolae expression vector
                     /note="Leishmania tarentolae expression vector"
                     /db_xref="taxon:1029641"
     rRNA            1..686
                     /product="18S ribosomal RNA"
                     /label=5 SSU 18S rRNA
                     /note="5 SSU 18S rRNA"
     misc_feature    1094..1096
                     /label=start codon
                     /note="start codon"
     misc_feature    1106..1123
                     /label=6xHis tag
                     /note="6xHis tag"
     CDS             1106..1123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1133..2116
                     /label=infrared fluorescent protein
                     /note="infrared fluorescent protein"
     CDS             1133..2116
                     /codon_start=1
                     /product="bacteriophytochrome-based monomeric infrared 
                     fluorescent protein (Shu et al., 2009)"
                     /label=IFP1.4
                     /translation="MARDPLPFFPPLYLGGPEITTENCEREPIHIPGSIQPHGALLTAD
                     GHSGEVLQVSLNAATFLGQEPTVLRGQTLAALLPEQWPALQAALPPGCPDALQYRATLD
                     WPAAGHLSLTVHRVAELLILEFEPTEAWDSIGPHALRNAMFALESAPNLRALAEVATQT
                     VRELTGFDRVMLYKFAPDATGEMIAEARREGMQAFLGHRFPASHTPAQARALYTRHLLR
                     LTADTRAAAVPLDPVLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGG
                     QLWGLIVCHHQTPYVLPPDLRTTLEELGRKLSGQVQRKEAGMDELYK"
     misc_feature    2117..2137
                     /label=TEV recognition site
                     /note="TEV recognition site"
     CDS             2117..2137
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    2156..2163
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    2164..2830
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    2831..2838
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             4209..4733
                     /codon_start=1
                     /product="nourseothricin resistance marker"
                     /label=nourseothricin resistance marker
                     /protein_id="AEE81079.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     rRNA            5357..6441
                     /product="18S ribosomal RNA"
                     /label=3 SSU 18S rRNA
                     /note="3 SSU 18S rRNA"
     promoter        complement(6459..6477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6498..6514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6522..6538
                     /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(6546..6576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6591..6612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6900..7488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7659..8519)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AEE81078.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7659..8519)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(8520..8624)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8651..9107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9248..9264
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9274..9292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"