LOCUS       sequence-36397-e         938 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 938)
  TITLE     2-series LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 938)
  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"
COMMENT     Sequence Label: pET GFP LIC cloning vector (2GFP-T)
FEATURES             Location/Qualifiers
     source          1..938
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(101..121)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(join(131..841,842..844))
                     /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(173..194)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     781..802
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(854..871)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             891..913
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ttcgggcttt gttagcagcc ggatcagcgc gcacgcgtgg taccggccgg ccaccaggtg
       61 gcgcgccgcg atcgcggatc cgttatccac ttccaatatt ggattggaag tacaggtttt
      121 cctcgatccc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
      181 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc
      241 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
      301 agtggtcggc gagctgcacg ctgccgtcct cgacgttgtg gcggatcttg aagttcgcct
      361 tgatgccgtt cttctgcttg tcggccgtga tatagacgtt gtggctgttg aagttgtact
      421 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
      481 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag gtgccgtcgt
      541 ccttgaagga gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
      601 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
      661 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
      721 gcttgccgtt ggtggcatcg ccctcgccct cgccgcgcac gctgaacttg tggccgttta
      781 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
      841 tcacagaaga accatggtga tggtgatggt gagaagattt catatgtata tctccttctt
      901 aaagttaaac aaaattattt ctagaactcg agtcctgc
//
