LOCUS       pETite_N-His_SUM        2535 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for Expresso(R) cloning to add N-terminal 6xHis and SUMO 
            tags, with inducible expression from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pETite N-His SUMO Kan
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2535)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 2535)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    21..42
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        51..69
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    71..86
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     RBS             113..135
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             143..145
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             146..163
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             176..463
                     /label=SUMO
                     /note="cleavable ubiquitin-like protein tag"
     terminator      495..542
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      640..669
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      691..1278
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1290..1317)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1344..1532)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(1536..2348)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(2349..2439)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(2440..2471)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
ORIGIN
        1 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc taatacgact
       61 cactataggg tgtgagcgga taacaatttc acgtggaaca gctagaaata attttgttta
      121 actataagaa ggagatatac atatgcatca tcaccaccat cacgggtccc tgcaggactc
      181 agaagtcaat caagaagcta agccagaggt caagccagaa gtcaagcctg agactcacat
      241 caatttaaag gtgtccgatg gatcttcaga gatcttcttc aagatcaaaa agaccactcc
      301 tttaagaagg ctgatggaag cgttcgctaa aagacagggt aaggaaatgg actccttaac
      361 gttcttgtac gacggtattg aaattcaagc tgatcagacc cctgaagatt tggacatgga
      421 ggataacgat attattgagg ctcaccgcga acagattgga ggttaataga gcggccgcca
      481 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      541 tgctgaaagg aggaactata tccgggtaac gaattcaagc ttgatatcat tcaggacgag
      601 cctcagactc cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc
      661 ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      781 accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      841 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      901 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      961 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1021 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1081 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1141 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1201 agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg
     1261 gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg
     1321 tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     1381 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     1441 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     1501 atgttaaggg cggttttttc ctgtttggtc atttagaaaa actcatcgag catcaagtga
     1561 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     1621 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     1681 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     1741 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
     1801 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     1861 gcaccaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     1921 ccgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     1981 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     2041 cctgggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     2101 gtcggaagag gcataaattc cgtcagccag tttagcctga ccatctcatc tgtaacatca
     2161 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     2221 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     2281 aaatcagcat ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata
     2341 tggctcatag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     2401 tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga
     2461 ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagta ttgagcgata
     2521 tctagagaat tcgtc
//
