LOCUS       sequence-287826-        2067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2067)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2067)
  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-m4046833
FEATURES             Location/Qualifiers
     source          1..2067
                     /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             450..470
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             471..488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      605..1193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1094..1113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1299..1958
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1365..1384)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2029..2063
                     /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 aacaatgaat
      121 tgagtccggt agcgctccgt caaatgtcgt gcgcagctgg gacgactcag acagcctgta
      181 ctgatgacaa cgctttagcc tattacaaca ccaccaaagg tggccgcttt gttctggctc
      241 tgctttccga cctgcaagat ctgaagtggg cgagattccc aaaaagcgac ggcaccggta
      301 ctatctacac ggagctggaa ccgccttgcc gattcgtcac cgatactccg aaaggtccca
      361 aggtgaaata cctatatttt attaaaggcc tgaataacct gaaccgcggc atggttctgg
      421 gttcactggc agctaccgtg cgtctgcaag agaacctgta ctttcaaagc caccatcatc
      481 accatcatta accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc
      541 gggtttttgc ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa
      601 gaaattgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaattgatcg
     1201 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     1261 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1321 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1381 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1441 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1501 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     1561 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1621 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1681 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtttcagcca
     1741 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1801 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     1861 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     1921 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     1981 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     2041 ggttgccgcc gggcgttttt tattggt
//
