LOCUS       sequence-295795-        7982 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 7982)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7982)
  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-m4133247
FEATURES             Location/Qualifiers
     source          1..7982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             878..898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             932..1009
                     /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             1037..1057
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1522..1643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1533..1552)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1587..1606
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1758..1804
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1834..1970
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2015..2086
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2312..2900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2392..2411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3400..4609
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4446..4465
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4613..5641
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5668..5716
                     /note="synthetic polyadenylation signal"
     CDS             join(5982..6050,6051..6842)
                     /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(6200..6219)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6855..6991
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7136..7207
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7212..7258)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7276..7324
                     /note="synthetic polyadenylation signal"
     enhancer        7405..7708
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7710..7912
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7863..7883
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc gccgttaagc ggacaatcaa agggacccat cactggttgc tccttactat
       61 actgacgtct ttactggtct tggtacagag tacacaatgg tcgcttttct tttttctgta
      121 cgagaatgca ttcctcccat ttgcgatggg aatcattgct atgtccgcct tcgctatgat
      181 gttcgtgaag cacaaacatg ccttcctttg cctgtttctc ctaccctcac tggcaaccgt
      241 ggcctatttt aacatggtgt acatgcctgc gagctgggtc atgcgaatta tgacttggct
      301 ggacatggtt gatacctccc tatctggctt caagttaaag gactgtgtga tgtatgcatc
      361 agctgtagtg ctgctaatcc tgatgacagc tcgcacagtc tacgatgacg gtgccaggag
      421 agtgtggact ctgatgaatg tcctcacctt agtttataaa gtatactatg gcaacgctct
      481 cgatcaggcc attagtatgt gggcattgat aatctccgtg acgagtaact acagcggagt
      541 ggtaaccact gtcatgtttt tggcacgcgg gatagtgttc atgtgtgtcg aatattgccc
      601 gatctttttt atcacaggga atactctgca atgcattatg ctggtttact gtttcttggg
      661 ctatttctgc acctgttatt ttggtctatt ctgcctgctt aataggtact ttcgtttgac
      721 actaggagtc tatgattact tggtgagcac gcaggagttt cggtatatga acagtcaggg
      781 cctgcttccc ccaaagaact ctattgacgc gtttaaactc aatataaagc tgttgggcgt
      841 gggtggaaaa ccttgcatca aggtggccac tgtgcaggaa aacctttact ttcagtcggg
      901 acggcgaatc ccggggctga taaatccctg gaaaaggaga tggaagaaga actttattgc
      961 agtgtctgcg gctaaccggt tcaaaaagat cagttcaagc ggcgccctag attacgacat
     1021 tcctacgacc gcctccgaga atctctattt ccagggtgaa tttggcttgg cacaacatga
     1081 cgaggccgtc gataacaagt ttaataaaga acagcagaac gctttctatg agatccttca
     1141 cttaccaaat ctgaacgaag agcaacgcaa cgctttcatt cagagcctga aagacgatcc
     1201 ctcgcagtct gcaaatctcc tggccgaagc caagaagctg aatgacgcac aggctccaaa
     1261 agttgataac aagtttaaca aggagcagca gaatgctttc tacgagatat tgcaccttcc
     1321 taatctgaat gaggaacaaa gaaatgcatt tatccagtcc ctaaaagacg atccatcaca
     1381 gagtgccaac ctgcttgcgg aagccaaaaa gctcaatgat gcccaggccc cgaaagtaga
     1441 cgcgaaccat caataaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt
     1501 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1561 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1621 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1681 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
     1741 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
     1801 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
     1861 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     1921 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
     1981 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
     2041 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
     2101 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
     2161 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
     2221 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
     2281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3061 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3121 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     3181 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     3241 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     3301 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     3361 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg
     3421 cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca
     3481 ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     3541 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     3601 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     3661 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     3721 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     3781 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     3841 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     3901 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     3961 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     4021 gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta
     4081 ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg
     4141 gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg
     4201 cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc
     4261 ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag
     4321 gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg
     4381 aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa
     4441 gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt
     4501 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac
     4561 ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa
     4621 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     4681 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     4741 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     4801 tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga
     4861 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     4921 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat
     4981 cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     5041 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     5101 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     5161 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     5221 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     5281 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     5341 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct
     5401 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     5461 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     5521 gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt
     5581 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     5641 agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
     5701 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
     5761 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     5821 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     5881 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     5941 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     6001 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6061 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     6121 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     6181 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6241 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     6301 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6361 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     6421 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6481 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6541 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     6601 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6661 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6721 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6781 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6841 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     6901 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     6961 cgtgtcctcc ttaccagaaa tttatcctta agatcgttta aactcgactc tggctctatc
     7021 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     7081 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     7141 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     7201 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     7261 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     7321 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     7381 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7441 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7501 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7561 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     7621 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     7681 ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg
     7741 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     7801 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     7861 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     7921 gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc
     7981 gc
//
