LOCUS       sequence-386453-        3545 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 3545)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3545)
  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-m4046841
FEATURES             Location/Qualifiers
     source          1..3545
                     /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..1958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      2079..2667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2568..2587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2773..3432
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2839..2858)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3503..3537
                     /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 aagcgactac aaggatgatg atgacaaata accggcttat cggtcagttt
     1981 cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat
     2041 gaatgactgt ccacgacgct atacccaaaa gaaaaagctt gagatccttt ttttctgcgc
     2101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2641 tcgtcagggg ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac
     2701 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     2761 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     2821 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     2881 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     2941 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca
     3001 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     3061 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     3121 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     3181 gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt
     3241 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     3301 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     3361 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     3421 ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac
     3481 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     3541 tcatc
//
