LOCUS       sequence-287902-        2088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF7a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2088)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2088)
  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-m4046901
FEATURES             Location/Qualifiers
     source          1..2088
                     /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             471..491
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             492..509
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      626..1214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1115..1134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1320..1979
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1386..1405)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2050..2084
                     /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 aagatcatac
      121 tattcctcgc cttgatcacc ctggcaacgt gtgaactgta tcactaccaa gagtgcgttc
      181 gcggtactac tgtcctgctt aaagaaccat gcagctcagg gacctacgaa ggcaactctc
      241 cgtttcatcc tctggcggac aataaattcg ctttaacatg cttttcgact cagttcgctt
      301 tcgcttgtcc cgatggcgta aaacacgtgt accagctgcg tgcgcgttcc gtttctccga
      361 aactgtttat tcgacaggag gaagtgcagg aactgtattc cccgattttc ctgatcgtag
      421 ccgcaatcgt tttcatcacc ctgtgcttta cccttaagcg caaaactgag gagaacctgt
      481 actttcaaag ccaccatcat caccatcatt aaccggctta tcggtcagtt tcacctgatt
      541 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg
      601 tccacgacgc tatacccaaa agaaattgag atcctttttt tctgcgcgta atctgctgct
      661 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      721 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      781 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      841 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      901 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      961 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1021 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1081 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1141 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1201 ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     1261 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     1321 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1381 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1441 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1501 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg
     1561 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1621 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1681 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1741 atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1801 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1861 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1921 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     1981 ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     2041 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggt
//
