LOCUS       sequence-287760-        1845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1845)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1845)
  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-m4046777
FEATURES             Location/Qualifiers
     source          1..1845
                     /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             222..242
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             243..266
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      383..971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     872..891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1077..1736
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1143..1162)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1807..1841
                     /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 ggctatataa
      121 acgttttcgc ttttccgttt acgatatata gtctactctt gtgcagaatg aattctcgta
      181 actacatagc acaagtagat gtagttaact ttaatctcac agagaacctg tactttcaaa
      241 gcgactacaa ggatgatgat gacaaataac cggcttatcg gtcagtttca cctgatttac
      301 gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc
      361 acgacgctat acccaaaaga aattgagatc ctttttttct gcgcgtaatc tgctgcttgc
      421 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      481 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      541 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      601 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      661 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      721 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      781 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      841 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      901 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      961 gcctatggaa attgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat
     1021 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     1081 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     1141 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     1201 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     1261 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     1321 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     1381 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     1441 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     1501 tgtttttcgt ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     1561 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     1621 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     1681 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg
     1741 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     1801 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggt
//
