LOCUS       sequence-22530-e         761 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 761)
  AUTHORS   Takahashi Y, Toh-E A, Kikuchi Y
  TITLE     Comparative analysis of yeast PIAS-type SUMO ligases in vivo and in 
            vitro.
  JOURNAL   J Biochem (Tokyo). 2003 Apr . 133(4):415-22.
  PUBMED    12761287
REFERENCE   2  (bases 1 to 761)
  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: "J Biochem 
            (Tokyo). 2003 Apr . 133(4):415-22."
COMMENT     Sequence Label: pT-43
FEATURES             Location/Qualifiers
     source          1..761
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     13..31
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(55..69,70..75,76..92,93..99)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    75..91
                     /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)."
     protein_bind    107..123
                     /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             142..153
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             172..189
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             196..489
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     terminator      527..621
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(731..750)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gaaatcataa
       61 aaaatttatt tgctttgtga gcggataaca attataatag attcaattgt gagcggataa
      121 caatttcaca cagaattcat taaagaggag aaattaacta tgagaggatc gcatcaccat
      181 caccatcacg gatccatgtc ggactcagaa gtcaatcaag aagctaagcc agaggtcaag
      241 ccagaagtca agcctgagac tcacatcaat ttaaaggtgt ccgatggatc ttcagagatc
      301 ttcttcaaga tcaaaaagac cactccttta agaaggctga tggaagcgtt cgctaaaaga
      361 cagggtaagg aaatggactc cttaagattc ttgtacgacg gtattagaat tcaagctgat
      421 cagacccctg aagatttgga catggaggat aacgatatta ttgaggctca cagagaacag
      481 attggtggtt agtcgacctg cagccaagct taattagctg agcttggact cctgttgata
      541 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
      601 gcgtttttta ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc
      661 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
      721 agaacatttt gaggcatttc agtcagttgc tcaatgtacc t
//
