LOCUS       sequence-295541-        6941 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 6941)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6941)
  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-m4134305
FEATURES             Location/Qualifiers
     source          1..6941
                     /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    481..602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(492..511)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     546..565
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      717..763
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      793..929
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    974..1045
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1271..1859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1351..1370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2359..3568
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3405..3424
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3572..4600
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4627..4675
                     /note="synthetic polyadenylation signal"
     CDS             join(4941..5009,5010..5801)
                     /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(5159..5178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5814..5950
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6095..6166
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6171..6217)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6235..6283
                     /note="synthetic polyadenylation signal"
     enhancer        6364..6667
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6669..6871
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6822..6842
                     /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 gttaaggatc
      421 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
      481 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      541 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      601 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      661 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
      721 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
      781 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
      841 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
      901 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
      961 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1021 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     1081 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     1141 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1201 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1681 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1801 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1861 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1981 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     2101 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     2161 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     2221 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     2281 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2341 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2401 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     2461 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     2521 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     2581 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     2641 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     2701 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     2761 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     2821 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     2881 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     2941 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3001 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     3061 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     3121 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3181 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3241 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3301 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3361 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3421 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     3481 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     3541 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     3601 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     3661 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     3721 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     3781 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     3841 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     3901 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     3961 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4021 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4081 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4141 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4201 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4261 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4321 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4381 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4441 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4501 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4561 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     4621 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     4681 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     4741 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     4801 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     4861 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     4921 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     4981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5041 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5101 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     5161 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5221 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     5281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5341 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5461 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     5521 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     5581 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5641 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5701 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5761 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     5821 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     5881 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     5941 ttatccttaa gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     6001 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     6061 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     6121 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     6181 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     6241 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     6301 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6361 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6421 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     6481 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     6541 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     6601 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6661 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     6721 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     6781 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     6841 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     6901 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
