LOCUS       sequence-296863-        2990 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 2990)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2990)
  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-m4046794
FEATURES             Location/Qualifiers
     source          1..2990
                     /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             1365..1385
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1386..1403
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1524..2112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2013..2032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2218..2877
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2284..2303)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2948..2982
                     /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 tcagataatg
      121 ggcctcaaaa ccagcgcaac gctccacgta taaccttcgg tggcccgtct gactccacag
      181 gcagcaacca gaatggtgag cgtagtggag ccaggtctaa gcagcgccgt ccccagggtc
      241 tgccgaacaa cacggcgagc tggtttactg cgcttaccca acacggtaaa gaagacttaa
      301 aattcccgcg tggccagggc gtcccgatca acactaactc ttccccggat gatcagattg
      361 gttattaccg cagagcaact cgtcggattc gcggcggcga cggtaaaatg aaagatttgt
      421 ctccgcgttg gtacttttac tatctcggca ccggtccgga agctggactg ccctacgggg
      481 caaacaagga cgggatcatt tgggtggcta ccgaaggtgc cctgaatact ccaaaagatc
      541 atatcggcac ccgtaaccct gctaataacg ctgcgatcgt tctgcaactg cctcagggta
      601 caactctgcc aaaaggcttc tatgcagaag ggtcgcgtgg tggcagccag gcgagtagcc
      661 gtagcagctc ccgctctcgc aactcgtccc gtaactccac tccgggctct tctcgcggca
      721 cgtccccggc tcgtatggca ggtaacggtg gtgacgctgc tctggcgctg ctactgctgg
      781 acaggctgaa ccagctggag tcgaagatga gtggtaaagg ccaacagcag cagggccaga
      841 ccgttaccaa aaagagcgca gctgaagcat ctaaaaaacc gcgacagaaa cgtaccgcga
      901 ctaaagcgta caatgtaaca caggccttcg gtcgtcgagg tccggagcaa actcaaggaa
      961 actttggcga tcaggaatta atccgccagg ggaccgacta caaacactgg ccgcaaattg
     1021 cccagttcgc cccatccgct agtgcattct tcggtatgag ccgcatcggc atggaagtaa
     1081 cgccaagtgg tacgtggctg acctacaccg gtgctatcaa gcttgatgac aaggacccga
     1141 actttaaaga ccaggttatt ctgcttaaca aacatattga tgcctataaa actttcccgc
     1201 cgaccgaacc caaaaaggac aaaaagaaaa aagctgatga aacccaggct ctaccgcagc
     1261 gtcagaaaaa acagcaaact gtgactctgc tgccagctgc ggacctggat gatttttcca
     1321 agcagctgca acagtctatg agctctgcgg actctacgca ggctgagaac ctgtactttc
     1381 aaagccacca tcatcaccat cattaaccgg cttatcggtc agtttcacct gatttacgta
     1441 aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg
     1501 acgctatacc caaaagaaaa agcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1561 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1621 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1981 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2041 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2101 agcctatgga aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     2161 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     2221 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     2281 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     2341 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     2401 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     2461 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     2521 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     2581 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     2641 atgtttttcg tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2701 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     2761 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2821 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     2881 gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct
     2941 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtcatc
//
