LOCUS       sequence-313543-        6966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6966)
  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-m4133209
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             919..939
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             943..1008
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1077..1198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1088..1107)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1142..1161
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1313..1359
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1389..1525
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1570..1641
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1867..2455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1947..1966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2955..4164
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4001..4020
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4168..5196
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5223..5271
                     /note="synthetic polyadenylation signal"
     CDS             join(5537..5605,5606..6397)
                     /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(5755..5774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6410..6546
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6698..6769
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6774..6820)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6838..6886
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaatga gcttagtcct
      601 gttgcactac gacagatgtc ttgtgctgcc ggtactacac aaactgcttg cactgatgac
      661 aatgcgttag cttactacaa cacaacaaag ggaggtaggt ttgtacttgc actgttatcc
      721 gatttacagg atttgaaatg ggctagattc cctaagagtg atggaactgg tactatctat
      781 acagaactgg aaccaccttg taggtttgtt acagacacac ctaaaggtcc taaagtgaag
      841 tatttatact ttattaaagg attaaacaac ctaaatagag gtatggtact tggtagttta
      901 gctgccacag tacgtctaga aaacctttac tttcagtcga tggattataa ggaccacgat
      961 ggagactaca aagatcatga cattgattac aaggacgacg atgacaaata aggatccgca
     1021 ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
     1081 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1141 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1201 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1261 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
     1321 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
     1381 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1441 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1501 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1561 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1621 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1681 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1741 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1801 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1861 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1921 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1981 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2041 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2101 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2161 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2221 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2281 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2341 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2401 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2461 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2521 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2701 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2761 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2821 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2881 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2941 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     3001 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     3061 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     3121 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     3181 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     3241 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     3301 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     3361 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     3421 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     3481 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     3541 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     3601 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     3661 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     3721 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     3781 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     3841 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     3901 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     3961 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     4021 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     4081 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     4141 ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct
     4201 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     4261 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     4321 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatccgcc
     4381 gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct gacctattgc
     4441 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     4501 gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct tagccagacg
     4561 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     4621 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     4681 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     4741 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     4801 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     4861 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     4921 cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct ccgcattggt
     4981 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     5041 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     5101 cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     5161 cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc ctagtcttag
     5221 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct
     5281 aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     5341 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     5401 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     5461 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     5521 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5581 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5641 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5701 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     5761 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5821 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     5881 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5941 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6001 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6061 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     6121 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     6181 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6241 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6301 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6361 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     6421 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     6481 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     6541 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     6601 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     6661 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     6721 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     6781 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     6841 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     6901 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6961 gttccg
//
