LOCUS       sequence-295043-        4587 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 4587)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4587)
  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-m4134428
FEATURES             Location/Qualifiers
     source          1..4587
                     /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             362..403
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    456..577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(467..486)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     521..540
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      692..738
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      768..904
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    949..1020
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1246..1834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1326..1345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2580..2648,2649..3440)
                     /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(2798..2817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3453..3589
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3741..3812
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3817..3863)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3881..3929
                     /note="synthetic polyadenylation signal"
     enhancer        4010..4313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4315..4517
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4468..4488
                     /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 ggggaaacct atacccaatc cacttctcgg cttagattct acctaagcta gcttgactga
      421 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
      481 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      541 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      601 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      661 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
      721 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
      781 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
      841 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
      901 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
      961 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     1021 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     1081 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     1141 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1201 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1261 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1321 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1381 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1441 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1501 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1561 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1621 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1681 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1741 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1801 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1861 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1921 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1981 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2041 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     2101 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     2161 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2221 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2281 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac
     2341 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     2401 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     2461 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     2521 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     2581 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2641 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     2701 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2761 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2821 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2881 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2941 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     3001 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3061 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3121 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     3181 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3241 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3301 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3361 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3421 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     3481 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     3541 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     3601 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     3661 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     3721 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     3781 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     3841 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     3901 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     3961 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4021 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4081 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4141 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4201 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4261 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     4321 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4381 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4441 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4501 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     4561 ccactcgaca cacccgccag cggccgc
//
