LOCUS       sequence-287829-        2157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2157)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2157)
  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-m4046968
FEATURES             Location/Qualifiers
     source          1..2157
                     /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             528..548
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             549..578
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      695..1283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1184..1203
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1389..2048
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1455..1474)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2119..2153
                     /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 gccggtaacg
      121 caacggaagt tcccgcgaat agcacagtgc tgtctttttg cgctttcgct gtagacgcgg
      181 caaaagctta caaggattat ttggcctcag gcggacagcc aattaccaac tgtgtcaaaa
      241 tgctctgcac tcacaccggg actggccaag ctataaccgt aactcctgag gcaaacatgg
      301 accaggaatc cttcggtggt gcgagttgct gcctgtactg tcgctgccat atcgatcacc
      361 cgaacccgaa aggcttttgt gacctgaagg ggaaatacgt tcagatcccg actacgtgcg
      421 ctaacgatcc ggttggtttc accctgaaaa acaccgtgtg caccgtgtgc ggtatgtgga
      481 aaggctatgg ctgttcttgt gaccagctga gggaaccgat gcttcaagag aacctgtact
      541 ttcaaagcga gcagaagctt atttccgaag aagatctcta accggcttat cggtcagttt
      601 cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat
      661 gaatgactgt ccacgacgct atacccaaaa gaaattgaga tccttttttt ctgcgcgtaa
      721 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      781 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      841 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      901 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      961 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1021 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1081 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1141 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1201 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1261 caggggggcg gagcctatgg aaattgatcg ggcacgtaag aggttccaac tttcaccata
     1321 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     1381 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1441 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1501 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1561 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga
     1621 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     1681 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     1741 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     1801 ttattgagaa tatgtttttc gtttcagcca atccctgggt gagtttcacc agttttgatt
     1861 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     1921 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1981 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2041 gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca
     2101 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggt
//
