LOCUS       sequence-287786-        3531 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 3531)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3531)
  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-m4046937
FEATURES             Location/Qualifiers
     source          1..3531
                     /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..1952
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      2069..2657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2558..2577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2763..3422
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2829..2848)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3493..3527
                     /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 ttcaaagcga ctacaaggat gatgatgaca aataaccggc ttatcggtca gtttcacctg
     1981 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
     2041 ctgtccacga cgctataccc aaaagaaatt gagatccttt ttttctgcgc gtaatctgct
     2101 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2161 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2221 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2281 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2341 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2401 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2461 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2521 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2581 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2641 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa
     2701 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     2761 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     2821 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     2881 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2941 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     3001 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     3061 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     3121 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     3181 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg
     3241 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     3301 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     3361 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     3421 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     3481 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg t
//
