LOCUS       sequence-297920-        6431 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 6431)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6431)
  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-m4134251
FEATURES             Location/Qualifiers
     source          1..6431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             525..545
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             549..614
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    667..788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(678..697)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     732..751
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      903..949
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      979..1115
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1160..1231
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1457..2045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1537..1556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2545..3754
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3591..3610
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3758..4357
                     /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(3758..3777)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4254..4274
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4384..4432
                     /note="synthetic polyadenylation signal"
     CDS             join(4698..4766,4767..5558)
                     /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(4916..4935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5571..5707
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5859..5930
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5935..5981)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5999..6047
                     /note="synthetic polyadenylation signal"
     enhancer        6128..6431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
      241 ctaccatcca ctcgacacac ccgccagcgg ccgccaccat gaaaatgtca gatgtaaagt
      301 gcacatcagt agtcttactc tcagttttgc aacaactcag agtagaatca tcatctaaat
      361 tgtgggctca atgtgtccag ttacacaatg acattctctt agctaaagat actactgaag
      421 cctttgaaaa aatggtttca ctactttctg ttttgctttc catgcagggt gctgtagaca
      481 taaacaagct ttgtgaagaa atgctggaca acagggcaac cttagaaaac ctttactttc
      541 agtcgatgga ttataaggac cacgatggag actacaaaga tcatgacatt gattacaagg
      601 acgacgatga caaataagct agcttgactg actgagatac agcgtacctt cagctcacag
      661 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      721 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      781 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
      841 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
      901 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
      961 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1021 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     1081 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     1141 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     1201 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     1261 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     1321 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     1381 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1441 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1501 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1561 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1621 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1681 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1741 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1801 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1861 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1921 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1981 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2041 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2101 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2161 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2221 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2281 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     2341 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     2401 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     2461 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     2521 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     2581 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     2641 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     2701 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     2761 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     2821 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     2881 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     2941 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     3001 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     3061 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     3121 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     3181 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     3241 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     3301 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     3361 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     3421 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     3481 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     3541 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     3601 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     3661 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     3721 taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt
     3781 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     3841 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     3901 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3961 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4021 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4081 gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4141 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4201 ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4261 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     4321 gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta
     4381 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4441 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     4501 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     4561 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     4621 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     4681 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4801 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4861 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     4921 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4981 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5101 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5221 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5281 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5521 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     5581 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     5641 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     5701 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     5761 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     5821 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     5881 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     5941 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     6001 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     6061 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6421 ctattaccat g
//
