LOCUS       sequence-295711-        7280 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 7280)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7280)
  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-m4133429
FEATURES             Location/Qualifiers
     source          1..7280
                     /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             710..727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    820..941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(831..850)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     885..904
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1056..1102
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1132..1268
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1313..1384
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1610..2198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1690..1709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2698..3907
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3744..3763
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3911..4939
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4966..5014
                     /note="synthetic polyadenylation signal"
     CDS             join(5280..5348,5349..6140)
                     /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(5498..5517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6153..6289
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6434..6505
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6510..6556)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6574..6622
                     /note="synthetic polyadenylation signal"
     enhancer        6703..7006
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7008..7210
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7161..7181
                     /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 gatgagtgac aaaatcattc atctaaccga tgactccttt gatacagacg tgttgaaggc
      421 cgatggtgcg atactggtag atttctgggc tgaatggtgt ggcccgtgca agatgatcgc
      481 acccattctg gacgagatcg ccgacgagta tcaaggaaaa ttaactgtcg ctaagcttaa
      541 tatagatcag aacccaggga cggcacctaa atacggcatc aggggaatac ctaccctcct
      601 gctctttaag aatggggaag ttgccgcgac aaaagtggga gccctgtcta agggccagct
      661 aaaagagttc ttggacgcta acctggcagg ttcaggctcg gggcacatgc accatcacca
      721 tcaccatagc agcgggctcg tgccacgcgg ctaaggatcc gcaggcctct gctagcttga
      781 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
      841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
      961 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1021 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
     1081 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
     1141 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     1201 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     1261 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     1321 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     1381 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     1441 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     1501 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     1561 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1621 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1681 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1801 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2041 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2341 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2401 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     2461 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     2521 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     2581 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     2641 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc
     2701 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca
     2761 cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     2821 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     2881 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     2941 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     3001 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     3061 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     3121 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     3181 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     3241 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     3301 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct
     3361 aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt
     3421 gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg
     3481 cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac
     3541 gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta
     3601 ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca
     3661 ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt
     3721 atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc
     3781 gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg
     3841 taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt
     3901 gttagacaac atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3961 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4021 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4081 tttctacaaa gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga
     4141 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4201 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4261 ggaggcaatg gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt
     4321 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4381 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4441 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4501 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4561 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4621 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4681 tgcaggatcg ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4741 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4801 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt
     4861 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4921 tccgagggca aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc
     4981 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     5041 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     5101 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     5161 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     5221 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     5281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     5401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5461 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5581 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     5701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5821 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     5881 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6121 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     6181 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     6241 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcgtttaaa
     6301 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     6361 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     6421 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     6481 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     6541 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     6601 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
     6661 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6721 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6781 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     6841 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     6901 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6961 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
     7021 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     7081 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     7141 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     7201 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
     7261 acacacccgc cagcggccgc
//
