LOCUS       sequence-110708-        1107 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  E coli expression plasmid to produce Thrombin cleavable His tagged 
            iLOV fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1107)
  TITLE     B Smith Lab iLOF fusion plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1107)
  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            "Unpublished"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: pET28-iLOVf
FEATURES             Location/Qualifiers
     source          1..1107
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(1..78)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     284..303
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     387..406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        387..405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    406..430
                     /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             445..467
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             474..488
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="MNHKV"
     CDS             849..866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             876..893
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             897..929
                     /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             978..995
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1048..1066)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
       61 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      121 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
      181 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
      241 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      301 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      361 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      421 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgaatc
      481 acaaagtgat gatcggtacc attgagaaga atttcgtaat cactgatcca cgtctgccgg
      541 acaacccgat catcttcgct tccgacggct tcctggagct gaccgagtat tcccgtgaag
      601 agatcctggg ccgtaacgcc cgtttcctgc agggcccgga gactgatcag gcgacggttc
      661 agaaaattcg cgatgctatc cgtgaccagc gtgaaactac ggtgcaactg attaactaca
      721 cgaaaagcgg taagaaattc tggaacctgc tgcatctgca accagtccgc gatcagaaag
      781 gcgagctgca gtatttcatc ggcgttcaac tggatggttc tgaccacgta cattccatgg
      841 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgg
      901 ctagcatgac tggtggacag caaatgggtc gggatccgaa ttcgagctcc gtcgacaagc
      961 ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc
     1021 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1081 cctctaaacg ggtcttgagg ggttttt
//
