LOCUS       sequence-298375-        3542 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 3542)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3542)
  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-m4046898
FEATURES             Location/Qualifiers
     source          1..3542
                     /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             1908..1928
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1929..1955
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      2076..2664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2565..2584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2770..3429
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2836..2855)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3500..3534
                     /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 gttggggctt
      121 gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt agaccattct
      181 tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta gtcttgtctg
      241 ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact caactttact
      301 taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt ccattgtgtg
      361 ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat aatgttactg
      421 actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt ttagctaaca
      481 cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact gaggagacat
      541 ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga gaattacatc
      601 tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc tttactggtt
      661 atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa aaaggtgact
      721 atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt ggtgattatt
      781 ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg ccacaagagc
      841 actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag ttttctagca
      901 atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag ggaccacctg
      961 gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct gctcgcatag
     1021 tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca ttaaaatatt
     1081 tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag tgttttgata
     1141 aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat gcattgcctg
     1201 agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat tatgatttga
     1261 gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac cctgctcaat
     1321 tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat ttcaattcag
     1381 tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt cggcgttgtc
     1441 ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt aaagcacata
     1501 aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg catgatgttt
     1561 catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca cgtaaccctg
     1621 cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta gcctcaaaga
     1681 ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat gactatgtca
     1741 tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt aatgttgcta
     1801 ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt tatgacaagt
     1861 tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttagag aacctgtact
     1921 ttcaaagcta cccgtatgac gttcctgatt atgcctaacc ggcttatcgg tcagtttcac
     1981 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
     2041 tgactgtcca cgacgctata cccaaaagaa aaagcttgag atcctttttt tctgcgcgta
     2101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2641 tcaggggggc ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat
     2701 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     2761 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     2821 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     2881 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2941 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     3001 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     3061 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     3121 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     3181 tttattgaga atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat
     3241 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     3301 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     3361 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     3421 ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc
     3481 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtca
     3541 tc
//
