LOCUS       sequence-292587-        2096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2096)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2096)
  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-m4046927
FEATURES             Location/Qualifiers
     source          1..2096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             433..453
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             454..471
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      592..1180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1081..1100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1286..1945
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1352..1371)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2016..2050
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     join(2078..2096,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2078..2096
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gactcttcta tgaaatttct ggtattctta ggtatcatta cgacagtggc tgccttccat
      121 caggagtgtt ctctgcaaag ctgcacccag caccagccat acgtcgttga tgacccgtgc
      181 cctatccact tctattccaa gtggtacatc cgagtaggcg cacgtaaaag cgcgccgttg
      241 attgaacttt gtgttgatga agcggggtcg aaaagtccca tacagtatat cgacattggc
      301 aattacactg tctcttgcct gccgtttact atcaactgcc aggaaccgaa gctcggttca
      361 ctggtggttc gctgctcttt ctacgaggac tttctggaat atcatgatgt tcgggttgtg
      421 ctggacttca tcgagaacct gtactttcaa agccaccatc atcaccatca ttaaccggct
      481 tatcggtcag tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga
      541 ggggcagaaa gatgaatgac tgtccacgac gctataccca aaagaaaaag cttgagatcc
      601 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      661 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      721 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      781 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      841 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      901 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      961 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1021 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1081 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1141 atttttgtga tgctcgtcag gggggcggag cctatggaaa ttgatcgggc acgtaagagg
     1201 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     1261 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     1321 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1381 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1441 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1501 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1561 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1621 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1681 ctatttccct aaagggttta ttgagaatat gtttttcgtt tcagccaatc cctgggtgag
     1741 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1801 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1861 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1921 cgatgagtgg cagggcgggg cgtaatttga tatcgagctc gcttggactc ctgttgatag
     1981 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
     2041 cgttttttat tggtcatcga tctcgatccc gcgaaattaa tacgactcac tatagg
//
