LOCUS       sequence-295472-        6496 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 6496)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6496)
  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-m4133880
FEATURES             Location/Qualifiers
     source          1..6496
                     /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             368..412
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    465..586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(476..495)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     530..549
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      701..747
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      777..913
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    958..1029
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1255..1843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1335..1354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2343..3552
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3389..3408
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3556..4155
                     /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(3556..3575)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4052..4072
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4182..4230
                     /note="synthetic polyadenylation signal"
     CDS             join(4496..4564,4565..5356)
                     /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(4714..4733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5369..5505
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5650..5721
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5726..5772)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5790..5838
                     /note="synthetic polyadenylation signal"
     enhancer        5919..6222
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6224..6426
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6377..6397
                     /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 tcagtcgaaa gaaaccgcgg cagctaagtt cgagaggcag cacatggaca gttaagctag
      421 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      481 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      541 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      601 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      661 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      721 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      781 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
      841 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
      901 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
      961 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1021 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1081 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1141 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2041 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     2101 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2161 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2221 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2281 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2341 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2401 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2461 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2521 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     2581 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     2641 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     2701 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     2761 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     2821 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     2881 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     2941 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     3001 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     3061 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     3121 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     3181 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     3241 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3301 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3361 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3421 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3481 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3541 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3601 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3661 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3721 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3781 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3841 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     3901 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3961 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     4021 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4081 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4141 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     4201 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4261 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4321 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4381 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4441 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4501 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4561 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     4621 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4681 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4741 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4801 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4861 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     4921 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4981 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5041 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     5101 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5161 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5221 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5281 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5341 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5401 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5461 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcg tttaaactcg
     5521 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     5581 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     5641 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     5701 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     5761 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     5821 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     5881 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     5941 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6001 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6061 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     6121 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6181 acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag
     6241 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     6301 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     6361 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     6421 agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac
     6481 acccgccagc ggccgc
//
