LOCUS       Exported                2659 bp ds-DNA    circular SYN 07-1-2015
DEFINITION  Vector for Expresso(R) cloning to add N-terminal 6xHis and SUMO tags
            and a C-terminal biotinylation tag, with rhamnose-inducible 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    pAviTag-C N-His SUMO Kan
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2659)
  AUTHORS   Lucigen
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..2659
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        1..119
                     /note="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)"
     RBS             153..175
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             183..185
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             186..203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             216..503
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="ubiquitin-like protein tag"
                     /note="SUMO"
                     /note="modified by Lucigen to prevent cleavage by 
                     eukaryotic SUMO proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLTFLYDGIEIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             543..587
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /note="AviTag(TM)"
                     /translation="GLNDIFEAQKIEWHE"
     terminator      619..666
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      764..793
                     /note="T3Te terminator"
                     /note="phage T3 early transcription terminator"
     rep_origin      815..1402
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      1414..1441
                     /note="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     CDS             complement(1465..1656)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1657..2472)
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2473..2563)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      2564..2595
                     /note="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"