LOCUS       Exported                2400 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046817
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2400)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2400)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2400
                     /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             774..794
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             795..821
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      938..1526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1427..1446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1632..2291
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1698..1717)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2362..2396
                     /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 gcagattcca
      121 acggtactat taccgttgaa gagcttaaaa agctccttga acaatggaac ctagtaatag
      181 gtttcctatt ccttacatgg atttgtcttc tacaatttgc ctatgccaac aggaataggt
      241 ttttgtatat aattaagtta attttcctct ggctgttatg gccagtaact ttagcttgtt
      301 ttgtgcttgc tgctgtttac agaataaatt ggatcaccgg tggaattgct atcgcaatgg
      361 cttgtcttgt aggcttgatg tggctcagct acttcattgc ttctttcaga ctgtttgcgc
      421 gtacgcgttc catgtggtca ttcaatccag aaactaacat tcttctcaac gtgccactcc
      481 atggcactat tctgaccaga ccgcttctag aaagtgaact cgtaatcgga gctgtgatcc
      541 ttcgtggaca tcttcgtatt gctggacacc atctaggacg ctgtgacatc aaggacctgc
      601 ctaaagaaat cactgttgct acatcacgaa cgctttctta ttacaaattg ggagcttcgc
      661 agcgtgtagc aggtgactca ggttttgctg catacagtcg ctacaggatt ggcaactata
      721 aattaaacac agaccattcc agtagcagtg acaatattgc tttgcttgta caggagaacc
      781 tgtactttca aagctacccg tatgacgttc ctgattatgc ctaaccggct tatcggtcag
      841 tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa
      901 gatgaatgac tgtccacgac gctataccca aaagaaattg agatcctttt tttctgcgcg
      961 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1021 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1081 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1141 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1201 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1261 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1321 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1381 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1441 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1501 cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc
     1561 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     1621 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     1681 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1741 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1801 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     1861 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1921 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1981 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     2041 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg
     2101 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     2161 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     2221 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     2281 gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc
     2341 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
