LOCUS       sequence-297357-        6437 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 6437)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6437)
  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-m4252813
FEATURES             Location/Qualifiers
     source          1..6437
                     /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             281..346
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    399..520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(410..429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     464..483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      635..681
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      711..847
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    892..963
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1189..1777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1269..1288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2277..3486
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3323..3342
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3490..4089
                     /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(3490..3509)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3986..4006
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4116..4164
                     /note="synthetic polyadenylation signal"
     CDS             join(4430..4498,4499..5290)
                     /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(4648..4667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5303..5439
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5591..5662
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5667..5713)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5731..5779
                     /note="synthetic polyadenylation signal"
     enhancer        5860..6163
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6165..6367
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6318..6338
                     /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 tcagtcgatg gattataagg accacgatgg
      301 agactacaaa gatcatgaca ttgattacaa ggacgacgat gacaaataag ctagcttgac
      361 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      421 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      481 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      541 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      601 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      661 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      721 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      781 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      841 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      901 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      961 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1021 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1081 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1141 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1201 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1261 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1321 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1381 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1441 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1501 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1561 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1621 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1681 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1741 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1801 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1861 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1921 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1981 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2041 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2101 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2161 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2221 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2281 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2341 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2401 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2461 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2521 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2581 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2641 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2701 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2761 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2821 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     2881 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     2941 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     3001 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     3061 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3121 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     3181 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3241 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3301 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3361 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3421 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3481 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3541 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3601 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3661 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3721 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3781 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3841 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3901 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     3961 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4021 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4081 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     4141 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4201 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4261 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4321 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4381 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4441 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4501 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4561 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4621 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4681 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4741 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4801 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4861 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4921 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4981 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5041 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5101 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5161 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5221 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5281 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5341 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5401 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5461 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5521 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5581 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5641 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5701 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5761 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5821 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5881 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5941 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6001 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6061 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6121 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     6181 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6241 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6301 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6361 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6421 cacccgccag cggccgc
//
