LOCUS       sequence-297851-        1862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1862)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1862)
  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-m4046953
FEATURES             Location/Qualifiers
     source          1..1862
                     /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             237..257
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             258..275
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      396..984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     885..904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1090..1749
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1156..1175)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1820..1854
                     /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 attgaattgt
      121 cactgatcga tttctatctt tgttttctcg cgttcctgct gtttctggtt ctgatcatgc
      181 tgattatctt ctggttctcc ttagagctgc aagaccataa tgaaacctgc cacgccgaga
      241 acctgtactt tcaaagccac catcatcacc atcattaacc ggcttatcgg tcagtttcac
      301 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
      361 tgactgtcca cgacgctata cccaaaagaa aaagcttgag atcctttttt tctgcgcgta
      421 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      481 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      541 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      601 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      661 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      721 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      781 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      841 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      901 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      961 tcaggggggc ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat
     1021 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     1081 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1141 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1201 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1261 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     1321 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1381 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1441 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1501 tttattgaga atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat
     1561 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1621 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1681 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1741 ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc
     1801 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtca
     1861 tc
//
