LOCUS       sequence-298369-        1991 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 1991)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1991)
  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-m4046832
FEATURES             Location/Qualifiers
     source          1..1991
                     /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..404
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      525..1113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1014..1033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1219..1878
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1285..1304)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1949..1983
                     /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 tggagccatc cgcagtttga aaaataaccg gcttatcggt
      421 cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag
      481 aaagatgaat gactgtccac gacgctatac ccaaaagaaa aagcttgaga tccttttttt
      541 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      601 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      661 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      721 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      781 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      841 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      901 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      961 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1021 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1081 tgatgctcgt caggggggcg gagcctatgg aaattgatcg ggcacgtaag aggttccaac
     1141 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     1201 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     1261 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     1321 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     1381 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt
     1441 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     1501 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     1561 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     1621 cctaaagggt ttattgagaa tatgtttttc gtttcagcca atccctgggt gagtttcacc
     1681 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     1741 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     1801 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     1861 tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt
     1921 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     1981 tattggtcat c
//
