LOCUS       sequence-307395-        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-m4046760
FEATURES             Location/Qualifiers
     source          1..3539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1786..1808
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1823..1847
                     /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)."
     primer_bind     complement(1847..1866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1848..1866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1894..1928)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1999..2658)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2573..2592
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(2764..3352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2844..2863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3473..3490)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3491..3511)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 atttcgaggc tcgtaaattg cagtttgtcg tacaggtcgc ggtcagacat gatgcagagg
       61 ataccaactt tggcgcgggt aatagctacg ttgaaccggt taacgttgca ggagtgagcg
      121 gtttcagtgg tttgggtgaa aatcacgtaa tcatattctg agccctggct agaatccaca
      181 gtctgcgtcg gcagacccag gattttggag gctactgcgt tctgagagtt atagggagag
      241 atgaagacag ctttacgcca tgccggattg cgggtgagaa attctcgcac aactccaatc
      301 tgtggacggt tgatcgcgct actgacgtcg tgcgtgataa cacctttata aaacattttg
      361 aagcactgag cggacttgtc cttatgggct ttcagtttgt tatcataaac cagagcagat
      421 acggtatcca caatttctgc cgggcatctg cgacacgtgc caagaaacat atctggaccg
      481 atcgtcttca tcagacggca aactgagttg aagtactccg gttccagggt gcctttagta
      541 agcagagtcc tcggtgccgg cagctgtgct gggtcgccga tgtatacgta gtgtttagca
      601 cgtaaacgcg cgttaactac gctcagatcg tagttggtcg ccatagagat ttcgtcgaac
      661 accacaatat cagcggtagt ttccggcagt gcattaacgg tacagaacac gtactgctcc
      721 agggtgctgt taactttaaa tttgtcaaag cattcaacac gagctctcgc cgggatgata
      781 cgagaacatt tatcaatagg gagatattta agggcctttt cgcacagtgc gtccaccgca
      841 gcgtgactgc atgcagtgta gacgatgcgc gcggatgggt aatacagggc cagacctata
      901 gcgaaatggc tttttccggt accaggcggg ccttgcagag tgctgtattt ttgcatacct
      961 actttctggt aattggctac gttagaggag aactcatccg agatgttcaa tgtcgggtac
     1021 agcccagtaa tgcgcacgta gtgttcctgc ggaaccagag tcggagcgga cagtggcatc
     1081 accgtgtgtg aagtcagaac gaagtagtca ccgacgttta acttatatgt agtggtgccc
     1141 cgatagacaa ctgcatcgcc gtagtcccct ttctcgaagg tgtattcacc gatctgcact
     1201 ttagagtttt ttgtaacacg atagccagta aaaacgtagt tgcggttcag aggcgggcgc
     1261 ggcttaccta cttcccagct cagatgcagt tcacgatcag atagaacctc acggaccgtt
     1321 gcaataccgt aggagagttt gaaggtttct tcggtcgcct tcagagtttc agcagcgaac
     1381 agtttcaatc gctcagtgca ggtattcgcc aggatatagt cgccagcgtt ggtccaatcg
     1441 caagttgcga tcgcgttaaa gtcagtcacg ttatcagacc caacacaggt atttttgtac
     1501 agaccgaata cctggccgtt ggcgcacaga ggaaaggata tcggtggttt gtggcttttg
     1561 caatagtaag acatgccacc caggtagagt tgagtcacgt ctgttacatc acatcccgga
     1621 gcgttgcaaa cgtacggatt aacggacaga accagcttgt gcgacgtact gattacatgg
     1681 tcataacagc atttgcaaca caggaaggga cgacgaatgc acgcgccgca gcgaaggctg
     1741 gtctgtgagt tgcataagac acatgcaccc acagccatag aagagtctcc ttcttaaagt
     1801 taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc
     1861 gtattaattt cgcgggatcg agatcgatga ccaataaaaa acgcccggcg gcaaccgagc
     1921 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     1981 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     2041 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     2101 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     2161 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     2221 tgaaacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     2281 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     2341 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     2401 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
     2461 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     2521 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     2581 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     2641 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     2701 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat
     2761 caatttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagctttttc
     3361 ttttgggtat agcgtcgtgg acagtcattc atctttctgc ccctccaaaa gcaaaaaccc
     3421 gccgaagcgg gtttttacgt aaatcaggtg aaactgaccg ataagccggt taatgatggt
     3481 gatgatggtg gctttgaaag tacaggttct cctgtaaagt agctacgtta cggcggggg
//
