LOCUS       sequence-66857-e         841 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 841)
  TITLE     Craig lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 841)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET15b/EYFP-GgVcl 884-1066 (Vt)
FEATURES             Location/Qualifiers
     source          1..841
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             16..38
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             57..74
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             84..101
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(105..107,108..110,111..821)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(150..171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(411..430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     758..779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc
       61 atcatcatca tcacagcagc ggcctggtgc cgcgcggcag ccatatggtg agcaagggcg
      121 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      181 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      241 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
      301 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      361 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      421 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      481 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      541 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
      601 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      661 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
      721 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      781 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtcgaattcg ggccccccct
      841 g
//
