LOCUS       pAviTag-N_N-His        2659 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for Expresso(R) cloning to add N-terminal 6xHis, 
            biotinylation, and SUMO tags, with rhamnose-inducible expression.
ACCESSION   .
VERSION     .
KEYWORDS    pAviTag-N N-His SUMO Kan.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2659)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 2659)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)"
     RBS             153..175
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             183..185
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             186..203
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             204..248
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     CDS             300..587
                     /label=SUMO
                     /note="cleavable ubiquitin-like protein tag"
     terminator      619..666
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      764..793
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      815..1402
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1414..1441)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1468..1656)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(1660..2472)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(2473..2563)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(2564..2595)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
ORIGIN
        1 caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc
       61 ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga ctggtcgtag
      121 ggagaccaca acggtttccc tctagaaata attttgttta actataagaa ggagatatac
      181 atatgcatca tcaccaccat cacggtctga acgacatctt cgaggctcag aaaatcgaat
      241 ggcacgaaag cttatccacc ccgccgaccc cgagcactcc tcctaccggg tccctgcagg
      301 actcagaagt caatcaagaa gctaagccag aggtcaagcc agaagtcaag cctgagactc
      361 acatcaattt aaaggtgtcc gatggatctt cagagatctt cttcaagatc aaaaagacca
      421 ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca gggtaaggaa atggactcct
      481 taacgttctt gtacgacggt attgaaattc aagctgatca gacccctgaa gatttggaca
      541 tggaggataa cgatattatt gaggctcacc gcgaacagat tggaggttaa tagagcggcc
      601 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      661 tttttgctga aaggaggaac tatatccggg taacgaattc aagcttgata tcattcagga
      721 cgagcctcag actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
      781 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      841 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      901 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
      961 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1141 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1201 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1321 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     1381 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     1441 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     1501 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     1561 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     1621 ggccatgtta agggcggttt tttcctgttt ggtcatttag aaaaactcat cgagcatcaa
     1681 gtgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1741 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1801 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1861 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1921 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1981 actcgcacca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2041 atcgccgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2101 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2161 tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2221 gatggtcgga agaggcataa attccgtcag ccagtttagc ctgaccatct catctgtaac
     2281 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2341 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2401 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     2461 aatatggctc atagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     2521 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     2581 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agtattgagc
     2641 gatatctaga gaattcgtc
//
