LOCUS       sequence-299796-        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-m4046949
FEATURES             Location/Qualifiers
     source          1..1964
                     /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             333..353
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             354..377
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      498..1086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     987..1006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1192..1851
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1258..1277)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1922..1956
                     /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 tactcattcg
      121 tttcggaaga gacaggtacg ttaatagtta atagcgtact tctttttctt gctttcgtgg
      181 tattcttgct agttacacta gccatcctta ctgcgcttcg attgtgtgcg tactgctgca
      241 atattgttaa cgtgagtctt gtaaaacctt ctttttacgt ttactctcgt gttaaaaatc
      301 tgaattcttc tagagttcct gatcttctgg tcgagaacct gtactttcaa agcgactaca
      361 aggatgatga tgacaaataa ccggcttatc ggtcagtttc acctgattta cgtaaaaacc
      421 cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc cacgacgcta
      481 tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      541 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      601 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      661 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      721 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      781 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      841 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      901 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      961 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1021 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1081 tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     1141 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa
     1201 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1261 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1321 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1381 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     1441 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1501 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1561 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1621 ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1681 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1741 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1801 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgatatc
     1861 gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
     1921 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt catc
//
