LOCUS       sequence-295424-        4629 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 4629)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4629)
  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-m4134259
FEATURES             Location/Qualifiers
     source          1..4629
                     /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..445
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    498..619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(509..528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     563..582
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      734..780
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      810..946
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    991..1062
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1288..1876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1368..1387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2622..2690,2691..3482)
                     /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(2840..2859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3495..3631
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3783..3854
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3859..3905)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3923..3971
                     /note="synthetic polyadenylation signal"
     enhancer        4052..4355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4357..4559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4510..4530
                     /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 tcagtcgaag aggcgatgga aaaagaattt tatcgcggtg agtgcagcca acagattcaa
      421 aaagatatcc tcgtctggtg ccctctaagc tagcttgact gactgagata cagcgtacct
      481 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      541 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      601 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
      661 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
      721 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      781 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
      841 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
      901 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
      961 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     1021 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     1081 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     1141 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     1201 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     1261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2101 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     2161 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     2221 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     2281 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     2341 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag
     2401 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     2461 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     2521 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     2581 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     2641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     2761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     2821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     2941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3001 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     3061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3181 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     3241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3481 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     3541 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     3601 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     3661 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     3721 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     3781 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     3841 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     3901 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     3961 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     4021 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4081 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4141 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4201 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4261 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4321 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     4381 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4441 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4501 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     4561 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     4621 agcggccgc
//
