LOCUS       sequence-295042-        7976 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 7976)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7976)
  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-m4134208
FEATURES             Location/Qualifiers
     source          1..7976
                     /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    1516..1637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1527..1546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1581..1600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1752..1798
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1828..1964
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2009..2080
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2306..2894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2386..2405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3394..4603
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4440..4459
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4607..5635
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5662..5710
                     /note="synthetic polyadenylation signal"
     CDS             join(5976..6044,6045..6836)
                     /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(6194..6213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6849..6985
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7130..7201
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7206..7252)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7270..7318
                     /note="synthetic polyadenylation signal"
     enhancer        7399..7702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7704..7906
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7857..7877
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7975..7976,1..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 ggatccgcag gcctctgcta gcttgactga ctgagataca gcgtaccttc
     1501 agctcacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1561 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1621 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1681 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc
     1741 tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
     1801 cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg
     1861 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     1921 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct
     1981 gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact
     2041 aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg
     2101 ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt
     2161 tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt
     2221 ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc
     2281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3061 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3121 ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct
     3181 ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg
     3241 ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat
     3301 atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg
     3361 agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc
     3421 ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg
     3481 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     3541 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     3601 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     3661 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     3721 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     3781 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     3841 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     3901 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     3961 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
     4021 tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg
     4081 tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     4141 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     4201 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     4261 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
     4321 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     4381 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     4441 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     4501 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa
     4561 attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc
     4621 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     4681 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     4741 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     4801 tcggcacttt gcatccgccg cgctcccgat tccggaagtg cttgacattg gggagttcag
     4861 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     4921 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc
     4981 cgccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     5041 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     5101 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     5161 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     5221 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     5281 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     5341 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc
     5401 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     5461 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     5521 cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg accgatggct gtgtagaagt
     5581 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataagctag
     5641 tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt
     5701 tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     5761 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     5821 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     5881 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     5941 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     6001 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6061 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     6121 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     6181 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6241 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     6301 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6361 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     6421 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6481 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     6541 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     6601 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6661 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6721 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6781 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     6841 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     6901 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     6961 ctccttacca gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct
     7021 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
     7081 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
     7141 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
     7201 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
     7261 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
     7321 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
     7381 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7441 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7501 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7561 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     7621 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     7681 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
     7741 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     7801 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     7861 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     7921 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
