LOCUS       sequence-295053-        5154 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 5154)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5154)
  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-m4134180
FEATURES             Location/Qualifiers
     source          1..5154
                     /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             893..970
                     /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"
     polyA_signal    1023..1144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1034..1053)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1088..1107
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1259..1305
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1335..1471
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1516..1587
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1813..2401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1893..1912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3147..3215,3216..4007)
                     /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(3365..3384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4020..4156
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4308..4379
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4384..4430)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4448..4496
                     /note="synthetic polyadenylation signal"
     enhancer        4577..4880
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4882..5084
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5035..5055
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5153..5154,1..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 cgaagaggcg
      901 atggaaaaag aattttatcg cggtgagtgc agccaacaga ttcaaaaaga tatcctcgtc
      961 tggtgccctc taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
     1021 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1081 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1141 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1201 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     1261 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     1321 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1381 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     1441 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     1501 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1561 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1621 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1681 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1741 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2221 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2641 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2701 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2761 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2821 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2881 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     2941 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     3001 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     3061 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     3121 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     3181 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3241 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     3301 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     3361 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3421 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     3481 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3541 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     3601 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3661 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     3721 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     3781 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3841 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3901 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3961 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     4021 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     4081 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     4141 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     4201 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     4261 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     4321 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     4381 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     4441 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     4501 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     4561 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4621 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4681 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4741 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4801 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4861 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4921 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4981 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     5041 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     5101 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
