LOCUS       Exported                1950 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046768
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1950)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1950
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      488..1076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     977..996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1182..1841
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1248..1267)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1912..1946
                     /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 agccaccatc
      361 atcaccatca ttaaccggct tatcggtcag tttcacctga tttacgtaaa aacccgcttc
      421 ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac gctataccca
      481 aaagaaattg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      601 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaattga
     1081 tcgggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     1141 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1201 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1261 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1321 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1381 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1441 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1501 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1561 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtttcag
     1621 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1681 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     1741 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     1801 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgatatc gagctcgctt
     1861 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     1921 ctcggttgcc gccgggcgtt ttttattggt
//
