LOCUS       sequence-292510-        2147 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 2147)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2147)
  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-m4046780
FEATURES             Location/Qualifiers
     source          1..2147
                     /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             526..546
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             547..564
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      685..1273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1174..1193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1379..2038
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1445..1464)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2109..2143
                     /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 tatgggtaat
      121 gcaacagaag tgcctgccaa ttcaactgta ttatctttct gtgcttttgc tgtagatgct
      181 gctaaagctt acaaagatta tctagctagt gggggacaac caatcactaa ttgtgttaag
      241 atgttgtgta cacacactgg tactggtcag gcaataacag ttacaccgga agccaatatg
      301 gatcaagaat cctttggtgg tgcatcgtgt tgtctgtact gccgttgcca catagatcat
      361 ccaaatccta aaggattttg tgacttaaaa ggtaagtatg tacaaatacc tacaacttgt
      421 gctaatgacc ctgtgggttt tacacttaaa aacacagtct gtaccgtctg cggtatgtgg
      481 aaaggttatg gctgtagttg tgatcaactc cgcgaaccca tgcttgagaa cctgtacttt
      541 caaagccacc atcatcacca tcattaaccg gcttatcggt cagtttcacc tgatttacgt
      601 aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag aaagatgaat gactgtccac
      661 gacgctatac ccaaaagaaa aagcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1261 gagcctatgg aaattgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag
     1321 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
     1381 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     1441 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     1501 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     1561 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
     1621 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1681 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     1741 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1801 tatgtttttc gtttcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1861 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     1921 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     1981 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     2041 tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     2101 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggt
//
