LOCUS       sequence-292508-        2330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2330)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2330)
  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-m4046852
FEATURES             Location/Qualifiers
     source          1..2330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             709..729
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             730..747
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      868..1456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1357..1376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1562..2221
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1628..1647)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2292..2326
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggccatt
      121 gcgagcgaat tttcaagtct tccatcctat gctgcattcg ctactgcaca agaggcttac
      181 gaacaggcgg tggccaacgg cgactcggaa gtcgttctga agaaactgaa aaaatcttta
      241 aatgttgcga aaagcgaatt tgatcgtgac gctgcgatgc agcgcaagtt ggagaaaatg
      301 gctgatcagg caatgacgca aatgtacaaa caggctcgtt ccgaagacaa aagggccaaa
      361 gtaacctctg ctatgcagac catgctgttc acaatgctgc gcaagctgga taacgacgcg
      421 ctcaacaaca tcatcaacaa tgcacgtgat ggttgcgtac ctctcaacat tatcccgctt
      481 actactgctg caaaactgat ggtggttata ccggactaca acacctataa gaacacctgt
      541 gatgggacga ctttcaccta cgccagcgca ctgtgggaaa tccagcaggt tgtcgatgct
      601 gacagtaaaa tcgttcagct gagcgagatt tctatggaca attctccgaa cctggcgtgg
      661 ccactgatcg taactgctct gcgtgcgaat agcgcggtca aattacaaga gaacctgtac
      721 tttcaaagcc accatcatca ccatcattaa ccggcttatc ggtcagtttc acctgattta
      781 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
      841 cacgacgcta tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg
      901 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      961 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1321 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1381 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1441 gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     1501 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1561 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1621 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1681 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1741 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     1801 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     1861 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     1921 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     1981 gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2041 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2101 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2161 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2221 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     2281 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
