LOCUS       sequence-287856-        1986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1986)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1986)
  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-m4046975
FEATURES             Location/Qualifiers
     source          1..1986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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             78..100
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             360..380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             381..407
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      524..1112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1013..1032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1218..1877
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1284..1303)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1948..1982
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg agcaaaatgt
      121 ctgatgtgaa gtgtacttcc gttgtattgc tgtcagtcct gcaacagctg cgtgttgagt
      181 cgtctagcaa actatgggca cagtgcgtgc agctgcacaa cgacatcctc ttagcgaaag
      241 atacgaccga agctttcgaa aaaatggttt cgctgttgtc cgtacttctg tctatgcaag
      301 gtgccgttga cattaataaa ctgtgcgaag agatgctgga caaccgcgct actctgcaag
      361 agaacctgta ctttcaaagc tacccgtatg acgttcctga ttatgcctaa ccggcttatc
      421 ggtcagtttc acctgattta cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg
      481 cagaaagatg aatgactgtc cacgacgcta tacccaaaag aaattgagat cctttttttc
      541 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      601 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      661 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      721 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      781 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      841 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      901 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      961 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     1021 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     1081 gatgctcgtc aggggggcgg agcctatgga aattgatcgg gcacgtaaga ggttccaact
     1141 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     1201 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     1261 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1321 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     1381 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta
     1441 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     1501 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     1561 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     1621 ctaaagggtt tattgagaat atgtttttcg tttcagccaa tccctgggtg agtttcacca
     1681 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     1741 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     1801 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     1861 ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta
     1921 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     1981 attggt
//
