LOCUS       sequence-299534-        6413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6413)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6413)
  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-m4258299
FEATURES             Location/Qualifiers
     source          1..6413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             278..322
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    375..496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(386..405)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     440..459
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      611..657
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      687..823
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    868..939
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1165..1753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1245..1264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2253..3462
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3299..3318
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3466..4065
                     /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(3466..3485)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3962..3982
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4092..4140
                     /note="synthetic polyadenylation signal"
     CDS             join(4406..4474,4475..5266)
                     /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(4624..4643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5279..5415
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5567..5638
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5643..5689)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5707..5755
                     /note="synthetic polyadenylation signal"
     enhancer        5836..6139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6141..6343
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6294..6314
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaagaaa acctttactt tcagtcgaaa gaaaccgcgg cagctaagtt
      301 cgagaggcag cacatggaca gttaagctag cttgactgac tgagatacag cgtaccttca
      361 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      421 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      481 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      541 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
      601 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
      661 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
      721 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
      781 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
      841 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
      901 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
      961 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1021 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1081 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1981 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2041 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2101 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2161 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2221 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     2281 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     2341 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     2401 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     2461 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     2521 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     2581 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     2641 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     2701 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     2761 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     2821 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     2881 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     2941 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     3001 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3061 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3121 gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3181 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     3241 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     3301 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     3361 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     3421 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     3481 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3541 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3601 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3661 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3721 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3781 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3841 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3901 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3961 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4021 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     4081 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     4141 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4201 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     4261 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4321 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     4381 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     4441 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4501 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     4561 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     4621 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4681 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     4741 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4801 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     4861 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4921 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     4981 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     5041 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5101 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5161 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5221 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     5281 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5341 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     5401 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     5461 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     5521 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     5581 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     5641 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     5701 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     5761 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     5821 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5881 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5941 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6001 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6061 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6121 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6181 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6241 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6301 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     6361 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
