LOCUS       sequence-295398-        6914 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 6914)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6914)
  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-m4134026
FEATURES             Location/Qualifiers
     source          1..6914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             347..367
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             368..385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    454..575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(465..484)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     519..538
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      690..736
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      766..902
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    947..1018
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1244..1832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1324..1343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2332..3541
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3378..3397
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3545..4573
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4600..4648
                     /note="synthetic polyadenylation signal"
     CDS             join(4914..4982,4983..5774)
                     /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(5132..5151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5787..5923
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6068..6139
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6144..6190)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6208..6256
                     /note="synthetic polyadenylation signal"
     enhancer        6337..6640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6642..6844
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6795..6815
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat aacgagttaa gcccagtagc gcttcgccag atgagttgcg ccgcaggtac
       61 aacccaaact gcttgtacgg atgacaacgc cctcgcttac tataatacca caaagggagg
      121 ccgattcgtt ctggccctgt tgtcggatct gcaagacttg aaatgggcac gttttcccaa
      181 gtccgacggg actggcacca tttacacaga actagagccg ccatgcaggt tcgtgactga
      241 tacccctaaa ggacccaagg tcaaatatct gtattttatc aaggggctta acaatctcaa
      301 tagaggcatg gtgttaggat ctctagccgc aacggtgcgg ctgcaagaaa acctttactt
      361 tcagtcgcac catcaccatc accattaagg atccgcaggc ctctgctagc ttgactgact
      421 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
      481 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      541 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      601 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
      661 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
      721 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
      781 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
      841 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
      901 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
      961 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     1021 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     1081 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     1141 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     1201 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1261 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1321 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1741 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1801 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2041 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     2101 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     2161 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     2221 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     2281 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     2341 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     2401 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     2461 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     2521 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     2581 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     2641 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     2701 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     2761 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     2821 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     2881 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     2941 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     3001 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     3061 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     3121 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     3181 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     3241 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     3301 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     3361 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     3421 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     3481 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     3541 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     3601 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     3661 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     3721 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     3781 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     3841 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     3901 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     3961 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4021 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4081 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4141 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4201 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4261 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4321 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4381 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4441 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     4501 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     4561 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     4621 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     4681 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     4741 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     4801 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     4861 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     4921 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4981 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     5041 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5101 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5161 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5221 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5281 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     5341 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5401 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5461 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     5521 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5581 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5641 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5701 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5761 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     5821 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     5881 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac
     5941 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     6001 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     6061 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     6121 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     6181 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     6241 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     6301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6601 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     6661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     6721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     6781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     6841 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     6901 ccgccagcgg ccgc
//
