LOCUS       sequence-292582-        1769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1769)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1769)
  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: pGBW-m4046974
FEATURES             Location/Qualifiers
     source          1..1769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      162..196
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     224..243
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        224..242
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    243..267
                     /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             282..304
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             348..368
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             369..386
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      507..1095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     996..1015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1201..1769,1..91)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1267..1286)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
       61 gtactgcgat gagtggcagg gcggggcgta atttgatatc gagctcgctt ggactcctgt
      121 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
      181 gccgggcgtt ttttattggt catcgatctc gatcccgcga aattaatacg actcactata
      241 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
      301 gagactcttc tatggctgat gcacaatcgt ttttaaacgg gtttgcggag aacctgtact
      361 ttcaaagcca ccatcatcac catcattaac cggcttatcg gtcagtttca cctgatttac
      421 gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc
      481 acgacgctat acccaaaaga aaaagcttga gatccttttt ttctgcgcgt aatctgctgc
      541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      601 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      661 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      721 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      781 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      841 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      901 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      961 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1021 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1081 cggagcctat ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata
     1141 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
     1201 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1261 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1321 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1381 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     1441 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1501 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1561 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1621 aatatgtttt tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1681 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     1741 gacaaggtgc tgatgccgct ggcgattca
//
