LOCUS       sequence-287836-        1959 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 1959)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1959)
  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-m4046730
FEATURES             Location/Qualifiers
     source          1..1959
                     /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..380
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      497..1085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     986..1005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1191..1850
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1257..1276)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1921..1955
                     /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 tatagttttg
      121 tatcagagga aacggggaca cttattgtca atagcgtgtt gctgttccta gctttcgttg
      181 tttttctgct cgtgacctta gcgatcctga ctgccttgcg tctgtgtgca tactgctgca
      241 acatcgttaa cgtatccctg gtaaagccgt ctttctacgt gtattcgcgc gttaaaaacc
      301 tcaacagctc tagggttccc gacctgcttg ttgagaacct gtactttcaa agctacccgt
      361 atgacgttcc tgattatgcc taaccggctt atcggtcagt ttcacctgat ttacgtaaaa
      421 acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact gtccacgacg
      481 ctatacccaa aagaaattga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      541 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      601 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      661 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      721 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      781 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      841 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      901 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      961 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1021 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1081 ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1141 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     1201 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1261 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     1321 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     1381 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     1441 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     1501 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     1561 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     1621 tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     1681 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     1741 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     1801 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg
     1861 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1921 ttcagaacgc tcggttgccg ccgggcgttt tttattggt
//
