LOCUS       sequence-300264-        2081 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 2081)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2081)
  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-m4046844
FEATURES             Location/Qualifiers
     source          1..2081
                     /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..494
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      615..1203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1104..1123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1309..1968
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1375..1394)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2039..2073
                     /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 gactacaagg
      481 atgatgatga caaataaccg gcttatcggt cagtttcacc tgatttacgt aaaaacccgc
      541 ttcggcgggt ttttgctttt ggaggggcag aaagatgaat gactgtccac gacgctatac
      601 ccaaaagaaa aagcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
      661 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      721 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      781 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      841 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      901 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      961 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1021 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1081 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1141 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1201 aaattgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     1261 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1321 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1381 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1441 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     1501 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     1561 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     1621 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1681 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1741 gtttcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1801 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1861 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1921 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag
     1981 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     2041 cagaacgctc ggttgccgcc gggcgttttt tattggtcat c
//
