LOCUS       sequence-99359-e         921 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pET28 driving expression of GFP, biobrick part Bba_E0040.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 921)
  AUTHORS   Shis DL, Bennett MR
  TITLE     Library of synthetic transcriptional AND gates built with split T7 
            RNA polymerase mutants.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5028-33. doi: 
            10.1073/pnas.1220157110. Epub 2013 Mar 11.
  PUBMED    23479654
REFERENCE   2  (bases 1 to 921)
  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: 
            "10.1073/pnas.1220157110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-03-26- 26"; volume: "110"; issue: "13"; pages: "5028-33"
COMMENT     Sequence Label: pET28:GFP
FEATURES             Location/Qualifiers
     source          1..921
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(19..36)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(83..796,797..799,800..802))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(86..796,797..799,800..802))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(128..149)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     477..496
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     736..757
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             809..831
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    846..870
                     /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(870..889)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(871..889)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gttagcagcc ggatctcagt ggtggtggtg gtggtgctcg agtgcggccg caagcttgtc
       61 gacggagctc gaattcggat ccttacttgt acagctcgtc catgccgaga gtgatcccgg
      121 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
      181 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
      241 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
      301 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
      361 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
      421 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
      481 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
      541 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
      601 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
      661 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
      721 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
      781 gctcctcgcc cttgctcacc atggtatatc tccttcttaa agttaaacaa aattatttct
      841 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
      901 tcgagatctc gatcctctac g
//
