LOCUS       sequence-292489-        2324 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 2324)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2324)
  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-m4046826
FEATURES             Location/Qualifiers
     source          1..2324
                     /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             703..723
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             724..741
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      862..1450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1351..1370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1556..2215
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1622..1641)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2286..2320
                     /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 tatgatagcc
      121 tcagagttta gttcccttcc atcatatgca gcttttgcta ctgctcaaga agcttatgag
      181 caggctgttg ctaatggtga ttctgaagtt gttcttaaaa agttgaagaa gtctttgaat
      241 gtggctaaat ctgaatttga ccgtgatgca gccatgcaac gtaagttgga aaagatggct
      301 gatcaagcta tgacccaaat gtataaacag gctagatctg aggacaagag ggcaaaagtt
      361 actagtgcta tgcagacaat gcttttcact atgcttagaa agttggataa tgatgcactc
      421 aacaacatta tcaacaatgc aagagatggt tgtgttccct tgaacataat acctcttaca
      481 acagcagcca aactaatggt tgtcatacca gactataaca catataaaaa tacgtgtgat
      541 ggtacaacat ttacttatgc atcagcattg tgggaaatcc aacaggttgt agatgcagat
      601 agtaaaattg ttcaacttag tgaaattagt atggacaatt cacctaattt agcatggcct
      661 cttattgtaa cagctttaag ggccaattct gctgtcaaat tagagaacct gtactttcaa
      721 agccaccatc atcaccatca ttaaccggct tatcggtcag tttcacctga tttacgtaaa
      781 aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac
      841 gctataccca aaagaaaaag cttgagatcc tttttttctg cgcgtaatct gctgcttgca
      901 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      961 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1021 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1081 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1141 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1201 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1261 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1321 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1381 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1441 cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg aaataagatc
     1501 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     1561 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     1621 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     1681 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     1741 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     1801 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     1861 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     1921 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     1981 gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     2041 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     2101 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     2161 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga
     2221 tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     2281 atttgttcag aacgctcggt tgccgccggg cgttttttat tggt
//
