LOCUS       sequence-313244-        6485 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 6485)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6485)
  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-m4134408
FEATURES             Location/Qualifiers
     source          1..6485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             925..945
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             946..972
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1025..1146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1036..1055)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1090..1109
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1261..1307
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1337..1473
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1518..1589
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1815..2403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1895..1914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2903..4112
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3949..3968
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4116..4715
                     /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(4116..4135)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4612..4632
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4742..4790
                     /note="synthetic polyadenylation signal"
     CDS             join(5056..5124,5125..5916)
                     /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(5274..5293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5929..6065
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6217..6288
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6293..6339)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6357..6405
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaataa cgagttaagc
      601 ccagtagcgc ttcgccagat gagttgcgcc gcaggtacaa cccaaactgc ttgtacggat
      661 gacaacgccc tcgcttacta taataccaca aagggaggcc gattcgttct ggccctgttg
      721 tcggatctgc aagacttgaa atgggcacgt tttcccaagt ccgacgggac tggcaccatt
      781 tacacagaac tagagccgcc atgcaggttc gtgactgata cccctaaagg acccaaggtc
      841 aaatatctgt attttatcaa ggggcttaac aatctcaata gaggcatggt gttaggatct
      901 ctagccgcaa cggtgcggct gcaagaaaac ctttactttc agtcgtaccc ctatgacgta
      961 ccagattatg cgtaagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     1021 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1081 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1141 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1201 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     1261 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     1321 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     1381 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     1441 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     1501 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     1561 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     1621 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     1681 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     1741 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     2641 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     2701 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     2761 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     2821 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     2881 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     2941 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
     3001 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     3061 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     3121 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     3181 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     3241 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     3301 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     3361 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     3421 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     3481 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     3541 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     3601 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3661 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     3721 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     3781 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     3841 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     3901 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     3961 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     4021 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
     4081 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
     4141 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4201 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4261 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4321 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4381 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4441 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4501 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4561 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4621 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4681 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
     4741 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     4801 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     4861 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     4921 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     4981 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     5041 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5101 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5161 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5221 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     5281 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5341 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     5401 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5461 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5521 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5581 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     5641 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     5701 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5761 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5821 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5881 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     5941 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     6001 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     6061 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6121 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     6181 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     6241 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     6301 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     6361 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     6421 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     6481 ttccg
//
