LOCUS       Exported                6349 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pDEST-LC2, complete sequence.
ACCESSION   JF327847
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  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 6349)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 6349)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6349
                     /organism="Expression vector LIC-pDEST-LC2"
                     /mol_type="other DNA"
                     /label=Escherichia coli expression vector
                     /note="Escherichia coli expression vector"
                     /db_xref="taxon:1029636"
     misc_feature    10..17
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    18..684
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    685..692
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    698..718
                     /label=TEV recognition site
                     /note="TEV recognition site"
     CDS             698..718
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    728..1711
                     /label=infrared fluorescent protein
                     /note="infrared fluorescent protein"
     CDS             728..1711
                     /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    1718..1738
                     /label=6xHis tag
                     /note="6xHis tag"
     CDS             1718..1735
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1736..1738
                     /label=stop codon (TAG)
                     /note="stop codon (TAG)"
     regulatory      1747..1875
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      1803..1850
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2216..2244)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2357..2461
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2462..3322
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AEE81071.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2462..3322
                     /gene="ampR"
                     /label=ampR
     rep_origin      3467..4140
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      3493..4081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4267..4409
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4511..4702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6267..6285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6267..6283
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     RBS             6317..6339
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      6332..6338
                     /regulatory_class="ribosome_binding_site"
     misc_feature    6347..6349
                     /label=start codon
                     /note="start codon"