LOCUS       sequence-64271-e         845 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 845)
  TITLE     Mutations_Koga_Nature
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 845)
  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            "Unpublished"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: FR52re
FEATURES             Location/Qualifiers
     source          1..845
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     69..88
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        69..87
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    88..112
                     /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)."
     RBS             127..149
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             157..402
                     /codon_start=1
                     /product="mechanically labile synthetic ferredoxin-like 
                     domain (Fang et al., 2013)"
                     /label=FL
                     /note="can serve as an intracellular tension sensor (Ringer
                     et al., 2017)"
                     /translation="MGEFDIRFRTDDDEQFEKVLKEMNRRARKDAGTVTYTRDGNDFEI
                     RITGISEQNRKELAKEVERLAKEQNITVTYTERGSLE"
     CDS             403..420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(473..491)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      487..534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(658..677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc
       61 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      121 aataattttg tttaacttta agaaggagat atacatatgg gtgaatttga tattcgcttc
      181 cgtaccgatg atgatgaaca gtttgaaaaa gtcctgaaag aaatgaaccg ccgtgctcgt
      241 aaagatgcgg gcaccgtgac gtatacccgt gatggtaacg actttgaaat tcgcatcacg
      301 ggcattagcg aacagaatcg taaagaactg gccaaagaag tggaacgcct ggcaaaagaa
      361 caaaacatta cggtcaccta cacggaacgc ggcagcctcg agcaccacca ccaccaccac
      421 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
      481 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
      541 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
      601 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      661 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      721 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      781 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      841 tgacg
//
