LOCUS       sequence-292506-        1958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1958)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1958)
  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-m4046768
FEATURES             Location/Qualifiers
     source          1..1958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      75..109
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     137..156
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        137..155
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    156..180
                     /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             195..217
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             450..470
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             471..488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      609..1197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1098..1117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1303..1958,1..4)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1369..1388)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtaatttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac
       61 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtcatcgat
      121 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
      181 cctctagaaa taattttgtt taactttaag aaggagactc ttctatgtac tcattcgttt
      241 cggaagagac aggtacgtta atagttaata gcgtacttct ttttcttgct ttcgtggtat
      301 tcttgctagt tacactagcc atccttactg cgcttcgatt gtgtgcgtac tgctgcaata
      361 ttgttaacgt gagtcttgta aaaccttctt tttacgttta ctctcgtgtt aaaaatctga
      421 attcttctag agttcctgat cttctggtcg agaacctgta ctttcaaagc caccatcatc
      481 accatcatta accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc
      541 gggtttttgc ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa
      601 gaaaaagctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg
     1201 atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     1261 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     1321 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     1381 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1441 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1501 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1561 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1621 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1681 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca
     1741 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1801 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     1861 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     1921 gaattacaac agtactgcga tgagtggcag ggcggggc
//
