LOCUS       sequence-295478-        7006 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 7006)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7006)
  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-m4133952
FEATURES             Location/Qualifiers
     source          1..7006
                     /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    975..1096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(986..1005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1040..1059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1211..1257
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1287..1423
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1468..1539
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1765..2353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1845..1864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2853..4062
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3899..3918
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4066..4665
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4066..4085)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4562..4582
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4692..4740
                     /note="synthetic polyadenylation signal"
     CDS             join(5006..5074,5075..5866)
                     /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(5224..5243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5879..6015
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6160..6231
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6236..6282)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6300..6348
                     /note="synthetic polyadenylation signal"
     enhancer        6429..6732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6734..6936
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6887..6907
                     /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 aataagctag cttgactgac tgagatacag cgtaccttca
      961 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1021 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1081 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1141 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     1201 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     1261 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     1321 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     1381 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     1441 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     1501 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     1561 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1621 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1681 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2581 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2641 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2701 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2761 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2821 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     2881 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     2941 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     3001 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     3061 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     3121 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     3181 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     3241 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     3301 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     3361 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     3421 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     3481 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     3541 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     3601 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3661 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3721 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3781 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     3841 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     3901 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     3961 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     4021 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     4081 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4141 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4201 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4261 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4321 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4381 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4441 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4501 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4561 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4621 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     4681 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     4741 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4801 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     4861 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4921 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     4981 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     5041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5101 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     5161 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     5221 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5281 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     5341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5401 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     5461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     5581 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     5641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     5881 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5941 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     6001 gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     6061 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     6121 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     6181 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     6241 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     6301 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     6361 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     6421 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6481 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6541 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6601 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6661 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6721 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6781 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6841 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6901 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     6961 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
