LOCUS       sequence-295625-        7451 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 7451)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7451)
  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-m4134324
FEATURES             Location/Qualifiers
     source          1..7451
                     /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             401..478
                     /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"
     CDS             506..526
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    991..1112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1002..1021)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1056..1075
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1227..1273
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1303..1439
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1484..1555
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1781..2369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1861..1880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2869..4078
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3915..3934
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4082..5110
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5137..5185
                     /note="synthetic polyadenylation signal"
     CDS             join(5451..5519,5520..6311)
                     /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(5669..5688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6324..6460
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6605..6676
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6681..6727)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6745..6793
                     /note="synthetic polyadenylation signal"
     enhancer        6874..7177
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7179..7381
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7332..7352
                     /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 tcagtcggga cggcgaatcc cggggctgat aaatccctgg aaaaggagat ggaagaagaa
      421 ctttattgca gtgtctgcgg ctaaccggtt caaaaagatc agttcaagcg gcgccctaga
      481 ttacgacatt cctacgaccg cctccgagaa tctctatttc cagggtgaat ttggcttggc
      541 acaacatgac gaggccgtcg ataacaagtt taataaagaa cagcagaacg ctttctatga
      601 gatccttcac ttaccaaatc tgaacgaaga gcaacgcaac gctttcattc agagcctgaa
      661 agacgatccc tcgcagtctg caaatctcct ggccgaagcc aagaagctga atgacgcaca
      721 ggctccaaaa gttgataaca agtttaacaa ggagcagcag aatgctttct acgagatatt
      781 gcaccttcct aatctgaatg aggaacaaag aaatgcattt atccagtccc taaaagacga
      841 tccatcacag agtgccaacc tgcttgcgga agccaaaaag ctcaatgatg cccaggcccc
      901 gaaagtagac gcgaaccatc aataaggatc cgcaggcctc tgctagcttg actgactgag
      961 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
     1021 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1081 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1141 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     1201 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
     1261 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
     1321 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     1381 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     1441 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
     1501 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
     1561 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     1621 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     1681 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     1741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2581 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     2641 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     2701 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     2761 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     2821 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
     2881 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
     2941 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     3001 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     3061 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     3121 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     3181 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     3241 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     3301 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     3361 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     3421 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     3481 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
     3541 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
     3601 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     3661 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     3721 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
     3781 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     3841 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     3901 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     3961 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     4021 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
     4081 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     4141 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     4201 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     4261 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga
     4321 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     4381 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat
     4441 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     4501 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     4561 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     4621 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     4681 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4741 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4801 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4861 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4921 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     4981 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga
     5041 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     5101 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga
     5161 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga
     5221 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     5281 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     5341 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     5401 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     5461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     5581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     5641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     5761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     5881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6001 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6061 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6121 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6181 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6241 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6301 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     6361 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     6421 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct
     6481 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     6541 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     6601 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     6661 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     6721 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     6781 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     6841 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     6901 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6961 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     7021 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     7081 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     7141 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     7201 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     7261 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     7321 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     7381 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     7441 ccagcggccg c
//
