LOCUS       sequence-295266-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5631)
  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-m4134037
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             872..892
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             926..1003
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1031..1051
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1500..1621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1511..1530)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1565..1584
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1736..1782
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1812..1948
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1993..2064
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2290..2878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2370..2389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3624..3692,3693..4484)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3842..3861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4497..4633
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4785..4856
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4861..4907)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4925..4973
                     /note="synthetic polyadenylation signal"
     enhancer        5054..5357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5359..5561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5512..5532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5630..5631,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gtgaaaagaa caatcaaggg tacacaccac tggttgttac tcacaatttt
       61 gacttcactt ttagttttag tccagagtac tcaatggtct ttgttctttt ttttgtatga
      121 aaatgccttt ttaccttttg ctatgggtat tattgctatg tctgcttttg caatgatgtt
      181 tgtcaaacat aagcatgcat ttctctgttt gtttttgtta ccttctcttg ccactgtagc
      241 ttattttaat atggtctata tgcctgctag ttgggtgatg cgtattatga catggttgga
      301 tatggttgat actagtttgt ctggttttaa gctaaaagac tgtgttatgt atgcatcagc
      361 tgtagtgtta ctaatcctta tgacagcaag aactgtgtat gatgatggtg ctaggagagt
      421 gtggacactt atgaatgtct tgacactcgt ttataaagtt tattatggta atgctttaga
      481 tcaagccatt tccatgtggg ctcttataat ctctgttact tctaactact caggtgtagt
      541 tacaactgtc atgtttttgg ccagaggtat tgtttttatg tgtgttgagt attgccctat
      601 tttcttcata actggtaata cacttcagtg tataatgcta gtttattgtt tcttaggcta
      661 tttttgtact tgttactttg gcctcttttg tttactcaac cgctacttta gactgactct
      721 tggtgtttat gattacttag tttctacaca ggagtttaga tatatgaatt cacagggact
      781 actcccaccc aagaatagca tagatgcctt caaactcaac attaaattgt tgggtgttgg
      841 tggcaaacct tgtatcaaag tagccactgt agaaaacctt tactttcagt cgggacggcg
      901 aatcccgggg ctgataaatc cctggaaaag gagatggaag aagaacttta ttgcagtgtc
      961 tgcggctaac cggttcaaaa agatcagttc aagcggcgcc ctagattacg acattcctac
     1021 gaccgcctcc gagaatctct atttccaggg tgaatttggc ttggcacaac atgacgaggc
     1081 cgtcgataac aagtttaata aagaacagca gaacgctttc tatgagatcc ttcacttacc
     1141 aaatctgaac gaagagcaac gcaacgcttt cattcagagc ctgaaagacg atccctcgca
     1201 gtctgcaaat ctcctggccg aagccaagaa gctgaatgac gcacaggctc caaaagttga
     1261 taacaagttt aacaaggagc agcagaatgc tttctacgag atattgcacc ttcctaatct
     1321 gaatgaggaa caaagaaatg catttatcca gtccctaaaa gacgatccat cacagagtgc
     1381 caacctgctt gcggaagcca aaaagctcaa tgatgcccag gccccgaaag tagacgcgaa
     1441 ccatcaataa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1501 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1561 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1621 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1681 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1741 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1801 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1861 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1921 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1981 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     2041 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     2101 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     2161 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     2221 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2281 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2341 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2401 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2461 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2521 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2581 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2641 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2701 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2761 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2821 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2881 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2941 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3001 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3061 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     3121 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     3181 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     3241 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     3301 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     3361 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     3421 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     3481 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     3541 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     3601 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     3661 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3721 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     3781 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     3841 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3901 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     3961 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4021 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     4081 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4141 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     4201 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     4261 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4321 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4381 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4441 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     4501 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     4561 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     4621 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     4681 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     4741 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     4801 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     4861 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     4921 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     4981 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     5041 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5101 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5161 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5221 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5281 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5341 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5401 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5461 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5521 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     5581 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
