LOCUS       sequence-295728-        7286 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 7286)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7286)
  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-m4134344
FEATURES             Location/Qualifiers
     source          1..7286
                     /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             716..733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    826..947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(837..856)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     891..910
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1062..1108
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1138..1274
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1319..1390
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1616..2204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1696..1715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2704..3913
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3750..3769
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3917..4945
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4972..5020
                     /note="synthetic polyadenylation signal"
     CDS             join(5286..5354,5355..6146)
                     /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(5504..5523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6159..6295
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6440..6511
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6516..6562)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6580..6628
                     /note="synthetic polyadenylation signal"
     enhancer        6709..7012
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7014..7216
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7167..7187
                     /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 tcagtcgatg agtgacaaaa tcattcatct aaccgatgac tcctttgata cagacgtgtt
      421 gaaggccgat ggtgcgatac tggtagattt ctgggctgaa tggtgtggcc cgtgcaagat
      481 gatcgcaccc attctggacg agatcgccga cgagtatcaa ggaaaattaa ctgtcgctaa
      541 gcttaatata gatcagaacc cagggacggc acctaaatac ggcatcaggg gaatacctac
      601 cctcctgctc tttaagaatg gggaagttgc cgcgacaaaa gtgggagccc tgtctaaggg
      661 ccagctaaaa gagttcttgg acgctaacct ggcaggttca ggctcggggc acatgcacca
      721 tcaccatcac catagcagcg ggctcgtgcc acgcggctaa ggatccgcag gcctctgcta
      781 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
      841 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      901 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      961 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1021 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
     1081 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
     1141 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
     1201 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
     1261 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
     1321 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     1381 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
     1441 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     1501 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     1561 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1621 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1681 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1741 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1801 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1861 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1921 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1981 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2041 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2101 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2161 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2221 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2281 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2341 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2401 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     2461 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     2521 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     2581 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     2641 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     2701 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     2761 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     2821 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2881 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     2941 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     3001 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     3061 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     3121 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     3181 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     3241 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     3301 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     3361 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     3421 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     3481 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     3541 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt
     3601 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     3661 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     3721 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     3781 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     3841 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     3901 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3961 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     4021 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     4081 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     4141 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     4201 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     4261 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     4321 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     4381 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     4441 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     4501 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4561 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4621 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4681 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4741 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4801 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4861 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     4921 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     4981 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     5041 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     5101 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     5161 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     5221 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     5281 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5341 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5401 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5461 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5521 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5581 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5641 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5701 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5761 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5821 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     5881 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     5941 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6001 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6061 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6121 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     6181 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     6241 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcg
     6301 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     6361 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     6421 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     6481 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     6541 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     6601 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     6661 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6721 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6781 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6841 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6901 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6961 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     7021 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     7081 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     7141 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     7201 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
     7261 cactcgacac acccgccagc ggccgc
//
