LOCUS       sequence-295002-        4935 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 4935)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4935)
  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-m4133355
FEATURES             Location/Qualifiers
     source          1..4935
                     /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"
     CDS             join(2928..2996,2997..3788)
                     /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(3146..3165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3801..3937
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4089..4160
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4165..4211)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4229..4277
                     /note="synthetic polyadenylation signal"
     enhancer        4358..4661
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4663..4865
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4816..4836
                     /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 tggtgtaccg agaacgatcc
     2701 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     2761 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     2821 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     2881 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     2941 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3001 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     3061 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     3121 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3181 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     3241 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3301 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     3361 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3421 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3481 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     3541 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3601 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3661 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3721 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3781 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     3841 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     3901 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     3961 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     4021 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     4081 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     4141 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     4201 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     4261 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     4321 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4381 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4441 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4501 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4561 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4621 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     4681 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4741 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4801 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4861 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     4921 cccgccagcg gccgc
//
