LOCUS       sequence-287803-        1854 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 1854)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1854)
  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-m4046859
FEATURES             Location/Qualifiers
     source          1..1854
                     /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      392..980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     881..900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1086..1745
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1152..1171)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1816..1850
                     /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 attgaacttt
      121 cattaattga cttctatttg tgctttttag cctttctgct attccttgtt ttaattatgc
      181 ttattatctt ttggttctca cttgaactgc aagatcataa tgaaacttgt cacgccgaga
      241 acctgtactt tcaaagccac catcatcacc atcattaacc ggcttatcgg tcagtttcac
      301 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
      361 tgactgtcca cgacgctata cccaaaagaa attgagatcc tttttttctg cgcgtaatct
      421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      961 gggggcggag cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg
     1021 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1081 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1141 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1201 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1261 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     1321 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1381 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1441 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1501 ttgagaatat gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1561 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1621 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     1681 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     1741 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     1801 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggt
//
