LOCUS       Exported                1026 bp ds-DNA     linear   SYN 17-MAY-2021
DEFINITION  Alive subunit for creating monovalent streptavidin with a single 
            femtomolar biotin binding site. Can also be used to create wild-type
            (tetravalent) streptavidin. .
ACCESSION   .
VERSION     .
KEYWORDS    pET21a-Streptavidin-Alive
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1026)
  AUTHORS   Howarth M, Chinnapen DJ, Gerrow K, Dorrestein PC, Grandy MR, 
            Kelleher NL, El-Husseini A, Ting AY
  TITLE     A monovalent streptavidin with a single femtomolar biotin binding 
            site.
  JOURNAL   Nat Methods. 2006 Apr . 3(4):267-73.
  PUBMED    16554831
REFERENCE   2  (bases 1 to 1026)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1026
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(40..57)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(85..102)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             494..516
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    531..555
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     complement(555..574)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(556..574)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(658..677)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        883..960
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(980..999)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctcagcttcc tttcgggctt tgttagcagc cggatctcag tggtggtggt ggtggtgctc
       61 gagtgcggcc gcaagctttt attaatggtg gtgatggtga tgggaagcag cggacggttt
      121 aactttggtg aaggtgtcgt gaccaaccag ggtggatttc cacgcgttgg cttcggtggt
      181 gccggaggtc aacaaccact gggtgttgat acgagcttca gcaccaccaa cgtactggcc
      241 agaccaggtg gtagcggagt gagcgttacg gtagttgttt ttccaagcaa cggtccaacc
      301 cagagcggtt ccggaaccgt cggtagccgg agcggagtcg taacgaccgg tcagaacgta
      361 tctagattca gcgttaccaa cagcggattc gtaggtaccg gtcagagcac cgtcagcacc
      421 agcggtaacg atgaaggtgg atcccagctg gttgtaccag gtgccggtga taccagcttc
      481 agccatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta
      541 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
      601 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
      661 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
      721 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
      781 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
      841 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
      901 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
      961 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     1021 tcccgc
//
