LOCUS       sequence-357431-        2999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 N (nucleocapsid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2999)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2999)
  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-m4046810
FEATURES             Location/Qualifiers
     source          1..2999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1346..1366
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1367..1393
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      1514..2102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2003..2022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2208..2867
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2274..2293)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2938..2972
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat gtcagataat gggcctcaaa accagcgcaa
      121 cgctccacgt ataaccttcg gtggcccgtc tgactccaca ggcagcaacc agaatggtga
      181 gcgtagtgga gccaggtcta agcagcgccg tccccagggt ctgccgaaca acacggcgag
      241 ctggtttact gcgcttaccc aacacggtaa agaagactta aaattcccgc gtggccaggg
      301 cgtcccgatc aacactaact cttccccgga tgatcagatt ggttattacc gcagagcaac
      361 tcgtcggatt cgcggcggcg acggtaaaat gaaagatttg tctccgcgtt ggtactttta
      421 ctatctcggc accggtccgg aagctggact gccctacggg gcaaacaagg acgggatcat
      481 ttgggtggct accgaaggtg ccctgaatac tccaaaagat catatcggca cccgtaaccc
      541 tgctaataac gctgcgatcg ttctgcaact gcctcagggt acaactctgc caaaaggctt
      601 ctatgcagaa gggtcgcgtg gtggcagcca ggcgagtagc cgtagcagct cccgctctcg
      661 caactcgtcc cgtaactcca ctccgggctc ttctcgcggc acgtccccgg ctcgtatggc
      721 aggtaacggt ggtgacgctg ctctggcgct gctactgctg gacaggctga accagctgga
      781 gtcgaagatg agtggtaaag gccaacagca gcagggccag accgttacca aaaagagcgc
      841 agctgaagca tctaaaaaac cgcgacagaa acgtaccgcg actaaagcgt acaatgtaac
      901 acaggccttc ggtcgtcgag gtccggagca aactcaagga aactttggcg atcaggaatt
      961 aatccgccag gggaccgact acaaacactg gccgcaaatt gcccagttcg ccccatccgc
     1021 tagtgcattc ttcggtatga gccgcatcgg catggaagta acgccaagtg gtacgtggct
     1081 gacctacacc ggtgctatca agcttgatga caaggacccg aactttaaag accaggttat
     1141 tctgcttaac aaacatattg atgcctataa aactttcccg ccgaccgaac ccaaaaagga
     1201 caaaaagaaa aaagctgatg aaacccaggc tctaccgcag cgtcagaaaa aacagcaaac
     1261 tgtgactctg ctgccagctg cggacctgga tgatttttcc aagcagctgc aacagtctat
     1321 gagctctgcg gactctacgc aggctgagaa cctgtacttt caaagctacc cgtatgacgt
     1381 tcctgattat gcctaaccgg cttatcggtc agtttcacct gatttacgta aaaacccgct
     1441 tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg acgctatacc
     1501 caaaagaaaa agcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1561 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1621 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1681 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1741 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1801 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1861 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1921 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1981 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2041 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2101 aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     2161 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     2221 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     2281 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     2341 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     2401 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     2461 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     2521 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     2581 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     2641 tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     2701 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2761 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     2821 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc
     2881 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     2941 agaacgctcg gttgccgccg ggcgtttttt attggtcatc gatctcgatc ccgcgaaat
//
