LOCUS       sequence-320327-        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-m4133344
FEATURES             Location/Qualifiers
     source          1..6437
                     /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             835..855
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             859..924
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    977..1098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(988..1007)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1042..1061
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1213..1259
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1289..1425
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1470..1541
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1767..2355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1847..1866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2855..4064
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3901..3920
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4068..4667
                     /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(4068..4087)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4564..4584
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4694..4742
                     /note="synthetic polyadenylation signal"
     CDS             join(5008..5076,5077..5868)
                     /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(5226..5245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5881..6017
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6169..6240
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6245..6291)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6309..6357
                     /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 ccatgagcaa aatgtctgac
      601 gtcaagtgta catccgtagt tctgttatca gtgcttcaac agctgcgggt ggagagttcg
      661 tcaaagctct gggcccagtg cgtgcagctg cacaatgata tattgctcgc gaaagatact
      721 acggaagctt tcgagaagat ggtcagcttg ctatctgttc tactgagcat gcaaggagca
      781 gtagacatca acaaactttg cgaagagatg ttagacaacc gagccaccct gcaagaaaac
      841 ctttactttc agtcgatgga ttataaggac cacgatggag actacaaaga tcatgacatt
      901 gattacaagg acgacgatga caaataagct agcttgactg actgagatac agcgtacctt
      961 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1021 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1081 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1141 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1201 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1261 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1321 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     1381 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     1441 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     1501 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     1561 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     1621 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     1681 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     1741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2341 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2461 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2521 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2581 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     2641 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     2701 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     2761 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     2821 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     2881 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     2941 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     3001 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     3061 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     3121 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3181 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     3241 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     3301 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     3361 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     3421 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     3481 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     3541 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     3601 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     3661 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     3721 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     3781 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     3841 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     3901 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     3961 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta
     4021 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca
     4081 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     4141 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4201 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4261 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4321 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4381 cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4441 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4501 tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4561 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4621 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag
     4681 cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
     4741 tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     4801 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     4861 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     4921 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     4981 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     5041 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5101 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     5161 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     5221 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5281 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     5341 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5401 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     5461 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5521 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     5581 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     5641 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5701 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5761 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5821 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     5881 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     5941 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     6001 cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc
     6061 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     6121 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6181 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6241 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6301 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6361 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6421 ccatatatgg agttccg
//
