LOCUS       pRham_N-His_SUMO        2575 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for Expresso(R) cloning to add N-terminal 6xHis and SUMO 
            tags, with rhamnose-inducible expression.
ACCESSION   .
VERSION     .
KEYWORDS    pRham N-His SUMO Kan
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2575)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 2575)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2575
                     /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             216..503
                     /label=SUMO
                     /note="cleavable ubiquitin-like protein tag"
     terminator      535..582
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      680..709
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      731..1318
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1330..1357)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1384..1572)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(1576..2388)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(2389..2479)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(2480..2511)
                     /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 cacgggtccc tgcaggactc agaagtcaat caagaagcta
      241 agccagaggt caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg
      301 gatcttcaga gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag
      361 cgttcgctaa aagacagggt aaggaaatgg actccttaac gttcttgtac gacggtattg
      421 aaattcaagc tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg
      481 ctcaccgcga acagattgga ggttaataga gcggccgcca ccgctgagca ataactagca
      541 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      601 tccgggtaac gaattcaagc ttgatatcat tcaggacgag cctcagactc cagcgtaact
      661 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
      721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
      841 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      901 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      961 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1021 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1081 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1141 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1201 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1261 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1321 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     1381 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     1441 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     1501 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     1561 ctgtttggtc atttagaaaa actcatcgag catcaagtga aactgcaatt tattcatatc
     1621 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1681 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1741 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1801 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg
     1861 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcaccaacca aaccgttatt
     1921 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ccgttaaaag gacaattaca
     1981 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2041 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag
     2101 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2161 cgtcagccag tttagcctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2221 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     2281 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2341 atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcatag ctcctgaaaa
     2401 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     2461 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
     2521 ggaggtcagg tatgatttaa atggtcagta ttgagcgata tctagagaat tcgtc
//
