LOCUS       sequence-120445-        1115 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pET-30-based vector dedicated to efficient scFv expression, which 
            circumvents the problem of in-frame amber codons in scFvs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1115)
  AUTHORS   Ossysek K, Uchanski T, Kulesza M, Bzowska M, Klaus T, Wos K, Madej 
            M, Bereta J
  TITLE     A new expression vector facilitating production and functional 
            analysis of scFv antibody fragments selected from Tomlinson I+J 
            phagemid libraries.
  JOURNAL   Immunol Lett. 2015 Jul 25. pii: S0165-2478(15)30011-0. doi: 
            10.1016/j.imlet.2015.07.005.
  PUBMED    26219832
REFERENCE   2  (bases 1 to 1115)
  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: "Immunol 
            Lett. 2015 Jul 25. pii: S0165-2478(15)30011-0. doi: 
            10.1016/j.imlet.2015.07.005."
COMMENT     Sequence Label: pET-scFv-T
FEATURES             Location/Qualifiers
     source          1..1115
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(2..79)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     285..304
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     392..411
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        392..410
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    411..435
                     /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             450..472
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     480..545
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             975..992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1020..1037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1090..1108)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
       61 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      121 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      181 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      241 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      301 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      361 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      421 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata
      481 tgaaatacct attgcctacg gcagccgctg gattgttatt actcgcggcc cagccggcca
      541 tggccgaggt gtttgactac tggggccagg gaaccctggt caccgtctcg agcggtggag
      601 gcggttcagg cggaggtggc agcggcggtg gcgggtcgac ggacatccag atgacccagt
      661 ctccatcctc cctgtctgca tctgtaggag acagagtcac catcacttgc cgggcaagtc
      721 agagcattag cagctattta aattggtatc agcagaaacc agggaaagcc cctaagctcc
      781 tgatctatcg ggcatccatg ttgcaaagtg gggtcccatc aaggttcagt ggcagtggat
      841 ctgggacaga tttcactctc accatcagca gtctgcaacc tgaagatttt gcaacttact
      901 actgtcaaca gattttgaag cggcctcata cgttcggcca agggaccaag gtggaaatca
      961 aacgggcggc cgcacatcat catcaccatc acaagcttta ataacgcgga tccttcgagc
     1021 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     1081 ctgctgccac cgctgagcaa taactagcat aaccc
//
