LOCUS       sequence-314727-        2070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 nsp3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2070)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2070)
  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-m4095418
FEATURES             Location/Qualifiers
     source          1..2070
                     /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             434..454
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             455..472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      589..1177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1078..1097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1283..1942
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1349..1368)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2013..2047
                     /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 gtttaccgaa cagcccatcg acttggtccc
      121 gaaccaacca tatcctaatg cgagcttcga taacttcaag tttgtatgcg acaacattaa
      181 attcgcagat gacttaaacc agctgacggg ttacaaaaaa ccggcttcgc gtgagcttaa
      241 ggttactttc ttcccggatc tgaacgggga cgtggttgcc atcgattaca aacactatac
      301 accttccttt aaaaaaggcg ctaagctgct acataaaccg attgtttggc acgtaaataa
      361 cgcgactaac aaagctacct acaaaccgaa cacttggtgt atccgctgcc tctggtctac
      421 taagccggtc gaagagaacc tgtactttca aagccaccat catcaccatc attaaccggc
      481 ttatcggtca gtttcacctg atttacgtaa aaacccgctt cggcgggttt ttgcttttgg
      541 aggggcagaa agatgaatga ctgtccacga cgctataccc aaaagaaatt gagatccttt
      601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      721 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1141 tttgtgatgc tcgtcagggg ggcggagcct atggaaattg atcgggcacg taagaggttc
     1201 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
     1261 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1321 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1381 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1441 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1501 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1561 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     1621 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1681 tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt
     1741 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1801 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1861 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1921 tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc
     1981 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     2041 tttttattgg tgatctcgat cccgcgaaat
//
