LOCUS       sequence-295914-        7445 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 7445)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7445)
  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-m4134376
FEATURES             Location/Qualifiers
     source          1..7445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             341..361
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             395..472
                     /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             500..520
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    985..1106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(996..1015)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1050..1069
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1221..1267
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1297..1433
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1478..1549
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1775..2363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1855..1874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2863..4072
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3909..3928
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4076..5104
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5131..5179
                     /note="synthetic polyadenylation signal"
     CDS             join(5445..5513,5514..6305)
                     /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(5663..5682)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6318..6454
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6599..6670
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6675..6721)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6739..6787
                     /note="synthetic polyadenylation signal"
     enhancer        6868..7171
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7173..7375
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7326..7346
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat gagcttagtc ctgttgcact acgacagatg tcttgtgctg ccggtactac
       61 acaaactgct tgcactgatg acaatgcgtt agcttactac aacacaacaa agggaggtag
      121 gtttgtactt gcactgttat ccgatttaca ggatttgaaa tgggctagat tccctaagag
      181 tgatggaact ggtactatct atacagaact ggaaccacct tgtaggtttg ttacagacac
      241 acctaaaggt cctaaagtga agtatttata ctttattaaa ggattaaaca acctaaatag
      301 aggtatggta cttggtagtt tagctgccac agtacgtcta gaaaaccttt actttcagtc
      361 gggacggcga atcccggggc tgataaatcc ctggaaaagg agatggaaga agaactttat
      421 tgcagtgtct gcggctaacc ggttcaaaaa gatcagttca agcggcgccc tagattacga
      481 cattcctacg accgcctccg agaatctcta tttccagggt gaatttggct tggcacaaca
      541 tgacgaggcc gtcgataaca agtttaataa agaacagcag aacgctttct atgagatcct
      601 tcacttacca aatctgaacg aagagcaacg caacgctttc attcagagcc tgaaagacga
      661 tccctcgcag tctgcaaatc tcctggccga agccaagaag ctgaatgacg cacaggctcc
      721 aaaagttgat aacaagttta acaaggagca gcagaatgct ttctacgaga tattgcacct
      781 tcctaatctg aatgaggaac aaagaaatgc atttatccag tccctaaaag acgatccatc
      841 acagagtgcc aacctgcttg cggaagccaa aaagctcaat gatgcccagg ccccgaaagt
      901 agacgcgaac catcaataag gatccgcagg cctctgctag cttgactgac tgagatacag
      961 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1021 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1081 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1141 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     1201 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1261 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     1321 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     1381 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     1441 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     1501 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     1561 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     1621 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     1681 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     1741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2581 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     2641 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     2701 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     2761 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2821 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2881 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2941 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     3001 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     3061 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     3121 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     3181 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3241 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3301 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3361 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3421 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3481 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     3541 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     3601 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     3661 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     3721 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     3781 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3841 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3901 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3961 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     4021 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     4081 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4141 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4201 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4261 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     4321 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4381 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     4441 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4501 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4561 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4621 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4681 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4741 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4801 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4861 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4921 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4981 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     5041 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     5101 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     5161 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     5221 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5281 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5341 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5401 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     5461 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5521 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     5581 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     5641 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5701 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5761 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5821 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5881 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5941 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6001 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     6061 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6121 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6181 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6241 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6301 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     6361 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     6421 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     6481 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     6541 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     6601 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     6661 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     6721 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     6781 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     6841 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6901 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6961 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7021 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     7081 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     7141 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     7201 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     7261 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7321 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     7381 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     7441 gccgc
//
