LOCUS       sequence-159520-         911 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of GFP nano body.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 911)
  AUTHORS   Kubala MH, Kovtun O, Alexandrov K, Collins BM
  TITLE     Structural and thermodynamic analysis of the GFP:GFP-nanobody 
            complex.
  JOURNAL   Protein Sci. 2010 Dec;19(12):2389-401. doi: 10.1002/pro.519.
  PUBMED    20945358
REFERENCE   2  (bases 1 to 911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/pro"; 
            journalName: "Protein Sci"; date: "2010-12"; volume: "19"; issue: 
            "12"; pages: "2389-401"
COMMENT     Sequence Label: pOPINE GFP nanobody
FEATURES             Location/Qualifiers
     source          1..911
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     88..107
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(197..214)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(218..559)
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid 
                     antibody (Rothbauer et al., 2008)"
                     /label=GFP nanobody
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGFEYW
                     GQGTQVTVSS"
     promoter        complement(580..689)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     protein_bind    705..729
                     /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(729..748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(730..748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(790..809)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(871..893)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct
       61 caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatcg atctcagtgg
      121 tatttgtgag ccagggcatt ggccacacca gccaccacct tctgataggc agcctgcacc
      181 tgaggttaat cacttagtga tggtgatggt gatgtttaga gctcaccgtc acctgagtcc
      241 cctggcccca atattcaaac cccacgttga cattgcaata atatactgcg gtgtcctccg
      301 gcttcagact gttcatttgc agatacacgg tattgcgcgc atcatcacgg ctaattgtaa
      361 aacgaccttt aacagagtcc tcgtaactag aacgatcgcc cgcactgctc atgccggcaa
      421 cccattcacg ttctttacca ggcgcctggc gataccaacg catagagtag cggttgactg
      481 ggaaaccgct tgcggcacag ctcaggcgca gactaccgcc cggttgtacc agagcaccgc
      541 cgctttccac cagttgaacc tgcatggtat atctcctttg attgtaaata aaatgtaatt
      601 tacagtatag tnttttaatt aatatacaaa tgatttgata ataattctta tttaactata
      661 atatattgtg ttgggttgaa ttaaaggtcc cggattaact ccggggaatt gttatccgct
      721 cacaattccc ctatagtgag tcgtattaat ttcggtccga tccaattctt tgccaaaatg
      781 atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga agaaggcatg
      841 aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga accccgccct
      901 gccccgtccc c
//
