LOCUS       sequence-287791-        3540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3540)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3540)
  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-m4046851
FEATURES             Location/Qualifiers
     source          1..3540
                     /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             1914..1934
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      2078..2666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2567..2586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2772..3431
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2838..2857)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3502..3536
                     /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 gctgtgggtg
      121 catgtgtctt atgcaactca cagaccagcc ttcgctgcgg cgcgtgcatt cgtcgtccct
      181 tcctgtgttg caaatgctgt tatgaccatg taatcagtac gtcgcacaag ctggttctgt
      241 ccgttaatcc gtacgtttgc aacgctccgg gatgtgatgt aacagacgtg actcaactct
      301 acctgggtgg catgtcttac tattgcaaaa gccacaaacc accgatatcc tttcctctgt
      361 gcgccaacgg ccaggtattc ggtctgtaca aaaatacctg tgttgggtct gataacgtga
      421 ctgactttaa cgcgatcgca acttgcgatt ggaccaacgc tggcgactat atcctggcga
      481 atacctgcac tgagcgattg aaactgttcg ctgctgaaac tctgaaggcg accgaagaaa
      541 ccttcaaact ctcctacggt attgcaacgg tccgtgaggt tctatctgat cgtgaactgc
      601 atctgagctg ggaagtaggt aagccgcgcc cgcctctgaa ccgcaactac gtttttactg
      661 gctatcgtgt tacaaaaaac tctaaagtgc agatcggtga atacaccttc gagaaagggg
      721 actacggcga tgcagttgtc tatcggggca ccactacata taagttaaac gtcggtgact
      781 acttcgttct gacttcacac acggtgatgc cactgtccgc tccgactctg gttccgcagg
      841 aacactacgt gcgcattact gggctgtacc cgacattgaa catctcggat gagttctcct
      901 ctaacgtagc caattaccag aaagtaggta tgcaaaaata cagcactctg caaggcccgc
      961 ctggtaccgg aaaaagccat ttcgctatag gtctggccct gtattaccca tccgcgcgca
     1021 tcgtctacac tgcatgcagt cacgctgcgg tggacgcact gtgcgaaaag gcccttaaat
     1081 atctccctat tgataaatgt tctcgtatca tcccggcgag agctcgtgtt gaatgctttg
     1141 acaaatttaa agttaacagc accctggagc agtacgtgtt ctgtaccgtt aatgcactgc
     1201 cggaaactac cgctgatatt gtggtgttcg acgaaatctc tatggcgacc aactacgatc
     1261 tgagcgtagt taacgcgcgt ttacgtgcta aacactacgt atacatcggc gacccagcac
     1321 agctgccggc accgaggact ctgcttacta aaggcaccct ggaaccggag tacttcaact
     1381 cagtttgccg tctgatgaag acgatcggtc cagatatgtt tcttggcacg tgtcgcagat
     1441 gcccggcaga aattgtggat accgtatctg ctctggttta tgataacaaa ctgaaagccc
     1501 ataaggacaa gtccgctcag tgcttcaaaa tgttttataa aggtgttatc acgcacgacg
     1561 tcagtagcgc gatcaaccgt ccacagattg gagttgtgcg agaatttctc acccgcaatc
     1621 cggcatggcg taaagctgtc ttcatctctc cctataactc tcagaacgca gtagcctcca
     1681 aaatcctggg tctgccgacg cagactgtgg attctagcca gggctcagaa tatgattacg
     1741 tgattttcac ccaaaccact gaaaccgctc actcctgcaa cgttaaccgg ttcaacgtag
     1801 ctattacccg cgccaaagtt ggtatcctct gcatcatgtc tgaccgcgac ctgtacgaca
     1861 aactgcaatt tacgagcctc gaaatccccc gccgtaacgt agctacttta caggagaacc
     1921 tgtactttca aagctacccg tatgacgttc ctgattatgc ctaaccggct tatcggtcag
     1981 tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa
     2041 gatgaatgac tgtccacgac gctataccca aaagaaattg agatcctttt tttctgcgcg
     2101 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2161 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2221 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2281 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2341 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2401 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2461 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2521 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2581 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2641 cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc
     2701 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     2761 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     2821 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     2881 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     2941 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     3001 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     3061 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     3121 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     3181 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg
     3241 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     3301 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     3361 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     3421 gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc
     3481 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
