LOCUS       sequence-313546-        1964 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 1964)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1964)
  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-m4046728
FEATURES             Location/Qualifiers
     source          1..1964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             314..334
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             335..358
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      479..1067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     968..987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1173..1832
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1239..1258)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1903..1937
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat gtactcattc gtttcggaag agacaggtac
      121 gttaatagtt aatagcgtac ttctttttct tgctttcgtg gtattcttgc tagttacact
      181 agccatcctt actgcgcttc gattgtgtgc gtactgctgc aatattgtta acgtgagtct
      241 tgtaaaacct tctttttacg tttactctcg tgttaaaaat ctgaattctt ctagagttcc
      301 tgatcttctg gtcgagaacc tgtactttca aagctggagc catccgcagt ttgaaaaata
      361 accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc gggtttttgc
      421 ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa gaaaaagctt
      481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg atcgggcacg
     1081 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     1141 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1201 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1261 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1321 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1381 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     1441 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     1501 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     1561 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct
     1621 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     1681 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     1741 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     1801 agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg
     1861 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1921 cgccgggcgt tttttattgg tcatcgatct cgatcccgcg aaat
//
