LOCUS       sequence-324350-        2416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2416)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2416)
  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-m4046980
FEATURES             Location/Qualifiers
     source          1..2416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             272..292
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             293..316
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        495..513
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    514..538
                     /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             553..575
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             766..786
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             787..810
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      931..1519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1420..1439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1625..2284
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1691..1710)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2355..2389
                     /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 gttccatctg gtcgactttc aagttactat
      121 tgctgaaata ttactgatca tcatgcgtac cttcaaggta tcgatctgga accttgatta
      181 cattatcaat ctcatcatta aaaacctgag taaatccctg acggagaaca aatattctca
      241 gttggatgaa gaacagccta tggagatcga cgagaacctg tactttcaaa gcgactacaa
      301 ggatgatgat gacaaataac cggcttatcg gtcagtttca cctgatttac gtaaaaaccc
      361 gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgctat
      421 acccaaaaga aaaagcttga gatccttttt ttctgcgcgt tttttattgg tcatcgatct
      481 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
      541 tctagaaata attttgttta actttaagaa ggagactctt ctatgttcca tctggtcgac
      601 tttcaagtta ctattgctga aatattactg atcatcatgc gtaccttcaa ggtatcgatc
      661 tggaaccttg attacattat caatctcatc attaaaaacc tgagtaaatc cctgacggag
      721 aacaaatatt ctcagttgga tgaagaacag cctatggaga tcgacgagaa cctgtacttt
      781 caaagcgact acaaggatga tgatgacaaa taaccggctt atcggtcagt ttcacctgat
      841 ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact
      901 gtccacgacg ctatacccaa aagaaaaagc ttgagatcct ttttttctgc gcgtaatctg
      961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1501 ggggcggagc ctatggaaat tgatcgggca cgtaagaggt tccaactttc accataatga
     1561 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     1621 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1681 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     1741 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1801 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga
     1861 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1921 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1981 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     2041 tgagaatatg tttttcgttt cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     2101 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     2161 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     2221 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     2281 gtaatttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac
     2341 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtcatcgat
     2401 ctcgatcccg cgaaat
//
