LOCUS       sequence-71640-e        1151 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 1151)
  TITLE     Series-9: N- or C-terminal Tag E.coli T7 Expression Vectors with 
            Biobrick Polycistronic Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1151)
  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            "Unpublished"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: pET His6 msfGFP TEV cloning vector with BioBri...
FEATURES             Location/Qualifiers
     source          1..1151
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     64..83
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        64..82
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             159..181
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             201..218
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(228..230,231..941)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(270..291)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     878..899
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             951..971
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1086..1104)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1100..1147
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
       61 aattaatacg actcactata gggagaccac aacggtttcc ctctagtgcc ggctccggag
      121 agctctttaa tgcggccgct taattaacta gaaataattt tgtttaactt taagaaggag
      181 atatacatat gaaatcttct caccatcacc atcaccatgg ttcttctgtg agcaagggcg
      241 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      301 acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
      361 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      421 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      481 agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctccttcaag gacgacggca
      541 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      601 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      661 tcaacagcca caacgtctat atcacggccg acaagcagaa gaacggcatc aaggcgaact
      721 tcaagatccg ccacaacgtc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      781 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      841 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      901 ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggggatcgag gaaaacctgt
      961 acttccaatc caatattgga agtggataac ggatcgcgat cgccctgcag gccggccggt
     1021 accacgcgtg cgcgctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1081 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1141 ttttttgctg a
//
