LOCUS       sequence-85159-e         994 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 488, Emission:
            507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 994)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 994)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: EGFP-pBAD
FEATURES             Location/Qualifiers
     source          1..994
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             77..99
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             119..136
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             140..172
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             185..205
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(212..214,215..217,218..931)
                     /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             join(212..214,215..217,218..928)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(257..278)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(518..537)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     865..886
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(977..994)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 ttagcggata ctactgacgc ttttatcgca actctctact gtttctccat acccgttttt
       61 ttgggctaga aataattttg tttaacttta agaaggagat atacatatgc ggggttctca
      121 tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct
      181 gtacgagaac ctgtacttcc agggctcgag catggtgagc aagggcgagg agctgttcac
      241 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      301 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      361 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      421 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      481 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      541 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      601 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      661 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
      721 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      781 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
      841 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      901 cactctcggc atggacgagc tgtacaagta agaattcgaa gcttggctgt tttggcggat
      961 gagagaagat tttcagcctg atacagatta aatc
//
