LOCUS       sequence-295484-        6493 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 6493)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6493)
  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-m4133903
FEATURES             Location/Qualifiers
     source          1..6493
                     /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..409
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    462..583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(473..492)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     527..546
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      698..744
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      774..910
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    955..1026
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1252..1840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1332..1351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2340..3549
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3386..3405
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3553..4152
                     /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(3553..3572)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4049..4069
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4179..4227
                     /note="synthetic polyadenylation signal"
     CDS             join(4493..4561,4562..5353)
                     /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(4711..4730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5366..5502
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5647..5718
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5723..5769)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5787..5835
                     /note="synthetic polyadenylation signal"
     enhancer        5916..6219
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6221..6423
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6374..6394
                     /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 tcagtcgggg aaacctatac ccaatccact tctcggctta gattctacct aagctagctt
      421 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      481 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      541 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      601 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      661 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
      721 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
      781 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
      841 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
      901 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
      961 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1021 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1081 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1141 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1201 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1261 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1321 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1381 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1441 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1501 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1561 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1621 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1681 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1741 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1801 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1921 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1981 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2041 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2101 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2161 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2221 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2281 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2341 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
     2401 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
     2461 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     2521 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     2581 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     2641 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     2701 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
     2761 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     2821 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     2881 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     2941 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
     3001 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
     3061 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     3121 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     3181 acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg
     3241 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     3301 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     3361 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     3421 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     3481 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     3541 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3601 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3661 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3721 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3781 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3841 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac
     3901 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3961 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg
     4021 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4081 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4141 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa
     4201 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac
     4261 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4321 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4381 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4441 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4501 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4561 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4621 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4681 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4741 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4801 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4861 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     4921 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4981 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5041 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     5101 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5161 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5221 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5281 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5341 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5401 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5461 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt aaactcgact
     5521 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     5581 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     5641 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     5701 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     5761 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
     5821 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
     5881 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5941 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     6001 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     6061 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     6121 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     6181 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
     6241 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     6301 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     6361 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     6421 gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc
     6481 cgccagcggc cgc
//
