LOCUS       sequence-295712-        6835 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 6835)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6835)
  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-m4133753
FEATURES             Location/Qualifiers
     source          1..6835
                     /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    804..925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(815..834)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     869..888
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1040..1086
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1116..1252
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1297..1368
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1594..2182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1674..1693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2682..3891
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3728..3747
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3895..4494
                     /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(3895..3914)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4391..4411
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4521..4569
                     /note="synthetic polyadenylation signal"
     CDS             join(4835..4903,4904..5695)
                     /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(5053..5072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5708..5844
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5989..6060
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6065..6111)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6129..6177
                     /note="synthetic polyadenylation signal"
     enhancer        6258..6561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6563..6765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6716..6736
                     /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 ctaagctagc ttgactgact gagatacagc
      781 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
      841 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
      901 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
      961 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1021 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1081 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1141 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1201 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1261 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1321 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1381 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1441 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1501 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2281 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2401 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2461 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2521 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2581 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2641 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     2701 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     2761 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     2821 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     2881 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     2941 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     3001 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     3061 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     3121 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     3181 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     3241 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     3301 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     3361 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     3421 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     3481 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     3541 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     3601 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     3661 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     3721 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     3781 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     3841 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     3901 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3961 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     4021 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     4081 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     4141 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     4201 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     4261 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4321 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     4381 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4441 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     4501 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     4561 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     4621 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     4681 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     4741 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     4801 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     4861 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4921 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     4981 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     5041 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5101 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     5161 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5221 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     5281 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5341 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     5401 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     5461 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5521 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5581 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5641 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     5701 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     5761 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     5821 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     5881 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     5941 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6001 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6061 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6121 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6181 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6241 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6301 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6361 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6421 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6481 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6541 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     6601 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     6661 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     6721 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     6781 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
