LOCUS       sequence-295707-        6974 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 6974)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6974)
  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-m4134298
FEATURES             Location/Qualifiers
     source          1..6974
                     /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..445
                     /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"
     polyA_signal    514..635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(525..544)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     579..598
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      750..796
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      826..962
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1007..1078
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1304..1892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1384..1403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2392..3601
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3438..3457
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3605..4633
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4660..4708
                     /note="synthetic polyadenylation signal"
     CDS             join(4974..5042,5043..5834)
                     /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(5192..5211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5847..5983
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6128..6199
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6204..6250)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6268..6316
                     /note="synthetic polyadenylation signal"
     enhancer        6397..6700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6702..6904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6855..6875
                     /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 tcagtcgaag aggcgatgga aaaagaattt tatcgcggtg agtgcagcca acagattcaa
      421 aaagatatcc tcgtctggtg ccctctaagg atccgcaggc ctctgctagc ttgactgact
      481 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
      541 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      601 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      661 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
      721 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
      781 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
      841 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
      901 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
      961 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     1021 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     1081 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     1141 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     1201 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     1261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2101 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     2161 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     2221 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     2281 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     2341 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     2401 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     2461 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     2521 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     2581 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     2641 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     2701 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     2761 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     2821 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     2881 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     2941 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     3001 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     3061 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     3121 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     3181 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     3241 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     3301 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     3361 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     3421 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     3481 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     3541 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     3601 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     3661 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     3721 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     3781 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     3841 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     3901 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     3961 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4021 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4081 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4141 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4201 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4261 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4321 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4381 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4441 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4501 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     4561 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     4621 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     4681 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     4741 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     4801 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     4861 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     4921 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     4981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     5101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5161 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5281 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     5401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5521 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     5581 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5821 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     5881 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     5941 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac
     6001 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     6061 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     6121 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     6181 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     6241 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     6301 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     6361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6661 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     6721 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     6781 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     6841 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     6901 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     6961 ccgccagcgg ccgc
//
