LOCUS       sequence-295677-        7817 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 7817)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7817)
  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-m4133267
FEATURES             Location/Qualifiers
     source          1..7817
                     /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             1247..1264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1357..1478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1368..1387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1422..1441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1593..1639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1669..1805
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1850..1921
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2147..2735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2227..2246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3235..4444
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4281..4300
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4448..5476
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5503..5551
                     /note="synthetic polyadenylation signal"
     CDS             join(5817..5885,5886..6677)
                     /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(6035..6054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6690..6826
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6971..7042
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7047..7093)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7111..7159
                     /note="synthetic polyadenylation signal"
     enhancer        7240..7543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7545..7747
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7698..7718
                     /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 ttcagtcgat
      901 gagtgacaaa atcattcatc taaccgatga ctcctttgat acagacgtgt tgaaggccga
      961 tggtgcgata ctggtagatt tctgggctga atggtgtggc ccgtgcaaga tgatcgcacc
     1021 cattctggac gagatcgccg acgagtatca aggaaaatta actgtcgcta agcttaatat
     1081 agatcagaac ccagggacgg cacctaaata cggcatcagg ggaataccta ccctcctgct
     1141 ctttaagaat ggggaagttg ccgcgacaaa agtgggagcc ctgtctaagg gccagctaaa
     1201 agagttcttg gacgctaacc tggcaggttc aggctcgggg cacatgcacc atcaccatca
     1261 ccatagcagc gggctcgtgc cacgcggcta aggatccgca ggcctctgct agcttgactg
     1321 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac
     1381 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1441 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1501 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1561 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg
     1621 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag
     1681 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     1741 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     1801 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa
     1861 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct
     1921 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg
     1981 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg
     2041 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag
     2101 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2161 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2221 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2281 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2341 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2401 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2461 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2521 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2581 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2641 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2701 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2821 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2881 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2941 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct
     3001 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac
     3061 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca
     3121 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca
     3181 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc
     3241 cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt
     3301 cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
     3361 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
     3421 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
     3481 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
     3541 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
     3601 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
     3661 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
     3721 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
     3781 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
     3841 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat
     3901 gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa
     3961 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     4021 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     4081 acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc
     4141 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     4201 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     4261 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     4321 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     4381 ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     4441 agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     4501 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     4561 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     4621 ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt
     4681 gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     4741 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     4801 ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg
     4861 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     4921 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     4981 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     5041 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     5101 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     5161 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     5221 aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     5281 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     5341 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg
     5401 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     5461 gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat
     5521 cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga
     5581 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     5641 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     5701 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     5761 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     5821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     5941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6001 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6121 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     6241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6361 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     6421 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6661 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     6721 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     6781 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc gtttaaactc
     6841 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     6901 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     6961 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     7021 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     7081 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     7141 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     7201 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     7261 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     7321 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     7381 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     7441 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     7501 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     7561 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     7621 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     7681 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     7741 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     7801 cacccgccag cggccgc
//
