LOCUS       sequence-323991-        3539 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 3539)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3539)
  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-m4046813
FEATURES             Location/Qualifiers
     source          1..3539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1912..1932
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1933..1956
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      2077..2665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2566..2585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2771..3430
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2837..2856)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3501..3535
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggttggg
      121 gcttgtgttc tttgcaattc acagacttca ttaagatgtg gtgcttgcat acgtagacca
      181 ttcttatgtt gtaaatgctg ttacgaccat gtcatatcaa catcacataa attagtcttg
      241 tctgttaatc cgtatgtttg caatgctcca ggttgtgatg tcacagatgt gactcaactt
      301 tacttaggag gtatgagcta ttattgtaaa tcacataaac cacccattag ttttccattg
      361 tgtgctaatg gacaagtttt tggtttatat aaaaatacat gtgttggtag cgataatgtt
      421 actgacttta atgcaattgc aacatgtgac tggacaaatg ctggtgatta cattttagct
      481 aacacctgta ctgaaagact caagcttttt gcagcagaaa cgctcaaagc tactgaggag
      541 acatttaaac tgtcttatgg tattgctact gtacgtgaag tgctgtctga cagagaatta
      601 catctttcat gggaagttgg taaacctaga ccaccactta accgaaatta tgtctttact
      661 ggttatcgtg taactaaaaa cagtaaagta caaataggag agtacacctt tgaaaaaggt
      721 gactatggtg atgctgttgt ttaccgaggt acaacaactt acaaattaaa tgttggtgat
      781 tattttgtgc tgacatcaca tacagtaatg ccattaagtg cacctacact agtgccacaa
      841 gagcactatg ttagaattac tggcttatac ccaacactca atatctcaga tgagttttct
      901 agcaatgttg caaattatca aaaggttggt atgcaaaagt attctacact ccagggacca
      961 cctggtactg gtaagagtca ttttgctatt ggcctagctc tctactaccc ttctgctcgc
     1021 atagtgtata cagcttgctc tcatgccgct gttgatgcac tatgtgagaa ggcattaaaa
     1081 tatttgccta tagataaatg tagtagaatt atacctgcac gtgctcgtgt agagtgtttt
     1141 gataaattca aagtgaattc aacattagaa cagtatgtct tttgtactgt aaatgcattg
     1201 cctgagacga cagcagatat agttgtcttt gatgaaattt caatggccac aaattatgat
     1261 ttgagtgttg tcaatgccag attacgtgct aagcactatg tgtacattgg cgaccctgct
     1321 caattacctg caccacgcac attgctaact aagggcacac tagaaccaga atatttcaat
     1381 tcagtgtgta gacttatgaa aactataggt ccagacatgt tcctcggaac ttgtcggcgt
     1441 tgtcctgctg aaattgttga cactgtgagt gctttggttt atgataataa gcttaaagca
     1501 cataaagaca aatcagctca atgctttaaa atgttttata agggtgttat cacgcatgat
     1561 gtttcatctg caattaacag gccacaaata ggcgtggtaa gagaattcct tacacgtaac
     1621 cctgcttgga gaaaagctgt ctttatttca ccttataatt cacagaatgc tgtagcctca
     1681 aagattttgg gactaccaac tcaaactgtt gattcatcac agggctcaga atatgactat
     1741 gtcatattca ctcaaaccac tgaaacagct cactcttgta atgtaaacag atttaatgtt
     1801 gctattacca gagcaaaagt aggcatactt tgcataatgt ctgatagaga cctttatgac
     1861 aagttgcaat ttacaagtct tgaaattcca cgtaggaatg tggcaacttt agagaacctg
     1921 tactttcaaa gctggagcca tccgcagttt gaaaaataac cggcttatcg gtcagtttca
     1981 cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga
     2041 atgactgtcc acgacgctat acccaaaaga aaaagcttga gatccttttt ttctgcgcgt
     2101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2641 gtcagggggg cggagcctat ggaaattgat cgggcacgta agaggttcca actttcacca
     2701 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     2761 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     2821 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     2881 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     2941 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     3001 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     3061 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     3121 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     3181 gtttattgag aatatgtttt tcgtttcagc caatccctgg gtgagtttca ccagttttga
     3241 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     3301 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     3361 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     3421 cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct
     3481 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggt
//
