LOCUS       sequence-287934-        2592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2592)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2592)
  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-m4046942
FEATURES             Location/Qualifiers
     source          1..2592
                     /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             975..995
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             996..1013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1130..1718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1619..1638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1824..2483
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1890..1909)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2554..2588
                     /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 gcagtgaaaa
      121 gaacaatcaa gggtacacac cactggttgt tactcacaat tttgacttca cttttagttt
      181 tagtccagag tactcaatgg tctttgttct tttttttgta tgaaaatgcc tttttacctt
      241 ttgctatggg tattattgct atgtctgctt ttgcaatgat gtttgtcaaa cataagcatg
      301 catttctctg tttgtttttg ttaccttctc ttgccactgt agcttatttt aatatggtct
      361 atatgcctgc tagttgggtg atgcgtatta tgacatggtt ggatatggtt gatactagtt
      421 tgtctggttt taagctaaaa gactgtgtta tgtatgcatc agctgtagtg ttactaatcc
      481 ttatgacagc aagaactgtg tatgatgatg gtgctaggag agtgtggaca cttatgaatg
      541 tcttgacact cgtttataaa gtttattatg gtaatgcttt agatcaagcc atttccatgt
      601 gggctcttat aatctctgtt acttctaact actcaggtgt agttacaact gtcatgtttt
      661 tggccagagg tattgttttt atgtgtgttg agtattgccc tattttcttc ataactggta
      721 atacacttca gtgtataatg ctagtttatt gtttcttagg ctatttttgt acttgttact
      781 ttggcctctt ttgtttactc aaccgctact ttagactgac tcttggtgtt tatgattact
      841 tagtttctac acaggagttt agatatatga attcacaggg actactccca cccaagaata
      901 gcatagatgc cttcaaactc aacattaaat tgttgggtgt tggtggcaaa ccttgtatca
      961 aagtagccac tgtagagaac ctgtactttc aaagccacca tcatcaccat cattaaccgg
     1021 cttatcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt tttgcttttg
     1081 gaggggcaga aagatgaatg actgtccacg acgctatacc caaaagaaat tgagatcctt
     1141 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1201 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1261 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1321 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1381 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1441 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1501 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1561 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1621 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1681 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac gtaagaggtt
     1741 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1801 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     1861 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1921 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1981 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     2041 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     2101 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2161 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2221 atttccctaa agggtttatt gagaatatgt ttttcgtttc agccaatccc tgggtgagtt
     2281 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2341 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2401 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2461 atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct gttgatagat
     2521 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     2581 ttttttattg gt
//
