LOCUS       sequence-295841-        4596 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 4596)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4596)
  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-m4133936
FEATURES             Location/Qualifiers
     source          1..4596
                     /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"
     CDS             join(2589..2657,2658..3449)
                     /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(2807..2826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3462..3598
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3750..3821
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3826..3872)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3890..3938
                     /note="synthetic polyadenylation signal"
     enhancer        4019..4322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4324..4526
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4477..4497
                     /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 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2401 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2461 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     2521 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     2581 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2701 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2761 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2821 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2881 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3001 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3121 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3181 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3301 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3421 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3481 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     3541 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     3601 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     3661 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     3721 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     3781 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     3841 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     3901 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     3961 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4021 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4081 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4141 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4201 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4261 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4321 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4381 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4441 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4501 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     4561 tcctaccatc cactcgacac acccgccagc ggccgc
//
