LOCUS       sequence-295624-        6841 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 6841)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6841)
  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-m4134362
FEATURES             Location/Qualifiers
     source          1..6841
                     /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             716..733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    810..931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(821..840)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     875..894
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1046..1092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1122..1258
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1303..1374
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1600..2188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1680..1699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2688..3897
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3734..3753
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3901..4500
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3901..3920)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4397..4417
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4527..4575
                     /note="synthetic polyadenylation signal"
     CDS             join(4841..4909,4910..5701)
                     /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(5059..5078)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5714..5850
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5995..6066
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6071..6117)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6135..6183
                     /note="synthetic polyadenylation signal"
     enhancer        6264..6567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6569..6771
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6722..6742
                     /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 tcagtcgatg agtgacaaaa tcattcatct aaccgatgac tcctttgata cagacgtgtt
      421 gaaggccgat ggtgcgatac tggtagattt ctgggctgaa tggtgtggcc cgtgcaagat
      481 gatcgcaccc attctggacg agatcgccga cgagtatcaa ggaaaattaa ctgtcgctaa
      541 gcttaatata gatcagaacc cagggacggc acctaaatac ggcatcaggg gaatacctac
      601 cctcctgctc tttaagaatg gggaagttgc cgcgacaaaa gtgggagccc tgtctaaggg
      661 ccagctaaaa gagttcttgg acgctaacct ggcaggttca ggctcggggc acatgcacca
      721 tcaccatcac catagcagcg ggctcgtgcc acgcggctaa gctagcttga ctgactgaga
      781 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      841 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      901 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      961 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1021 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1081 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1141 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1201 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1261 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1321 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1381 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1441 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1501 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1561 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1621 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1681 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1741 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1801 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1861 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1921 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1981 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2041 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2101 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2161 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2221 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2281 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2401 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2461 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2521 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2581 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2641 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg
     2701 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     2761 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     2821 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     2881 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     2941 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     3001 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     3061 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     3121 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     3181 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     3241 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     3301 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     3361 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     3421 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     3481 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     3541 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     3601 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     3661 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     3721 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     3781 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     3841 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     3901 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     3961 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     4021 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4081 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4141 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4201 tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag
     4261 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4321 agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg
     4381 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4441 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag
     4501 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     4561 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     4621 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     4681 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     4741 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     4801 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     4861 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4921 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     4981 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     5041 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5101 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     5161 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5221 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     5281 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5341 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5401 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     5461 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5521 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5581 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5641 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5701 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     5761 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     5821 gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct ggctctatcg
     5881 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     5941 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     6001 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     6061 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     6121 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     6181 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     6241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     6541 tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt
     6601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6721 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     6781 aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg
     6841 c
//
