LOCUS       sequence-295492-        6935 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 6935)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6935)
  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-m4133825
FEATURES             Location/Qualifiers
     source          1..6935
                     /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             362..406
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    475..596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(486..505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     540..559
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      711..757
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      787..923
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    968..1039
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1265..1853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1345..1364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2353..3562
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3399..3418
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3566..4594
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4621..4669
                     /note="synthetic polyadenylation signal"
     CDS             join(4935..5003,5004..5795)
                     /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(5153..5172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5808..5944
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6089..6160
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6165..6211)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6229..6277
                     /note="synthetic polyadenylation signal"
     enhancer        6358..6661
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6663..6865
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6816..6836
                     /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 gaaagaaacc gcggcagcta agttcgagag gcagcacatg gacagttaag gatccgcagg
      421 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      481 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      541 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      601 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      661 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      721 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      781 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      841 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      901 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
      961 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     1021 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     1081 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     1141 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     1201 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1261 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1321 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1381 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1441 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1501 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1561 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1621 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1681 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1741 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1801 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1861 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1921 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1981 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2041 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     2101 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     2161 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     2221 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     2281 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     2341 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2401 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2461 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2521 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2581 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2641 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2701 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2761 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2821 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2881 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     2941 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     3001 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     3061 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     3121 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     3181 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     3241 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3301 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3361 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3421 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3481 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3541 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     3601 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3661 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3721 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     3781 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     3841 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3901 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     3961 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4021 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4081 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4141 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4201 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4261 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4321 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4381 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4441 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4501 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4561 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     4621 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4681 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4741 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     4801 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4861 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4921 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4981 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5041 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5101 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5161 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5221 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     5281 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5341 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5401 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5461 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5521 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5581 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5641 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5701 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5761 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     5821 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5881 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     5941 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     6001 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     6061 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     6121 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     6181 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     6241 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     6301 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     6361 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     6421 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6481 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6541 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     6601 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6661 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     6721 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     6781 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     6841 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     6901 cctaccatcc actcgacaca cccgccagcg gccgc
//
