LOCUS       sequence-295223-        6397 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 6397)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6397)
  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-m4133324
FEATURES             Location/Qualifiers
     source          1..6397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             272..313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    366..487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(377..396)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     431..450
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      602..648
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      678..814
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    859..930
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1156..1744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1236..1255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2244..3453
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3290..3309
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3457..4056
                     /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(3457..3476)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3953..3973
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4083..4131
                     /note="synthetic polyadenylation signal"
     CDS             join(4397..4465,4466..5257)
                     /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(4615..4634)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5270..5406
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5551..5622
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5627..5673)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5691..5739
                     /note="synthetic polyadenylation signal"
     enhancer        5820..6123
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6125..6327
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6278..6298
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc ggggaaacct atacccaatc cacttctcgg
      301 cttagattct acctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
      361 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      421 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      481 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      541 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
      601 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
      661 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
      721 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
      781 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
      841 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
      901 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
      961 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     1021 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     1081 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1981 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     2041 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     2101 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     2161 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     2221 aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc
     2281 ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct
     2341 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     2401 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     2461 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     2521 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     2581 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     2641 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga
     2701 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct
     2761 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag
     2821 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa
     2881 gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc
     2941 tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     3001 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     3061 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     3121 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     3181 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     3241 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     3301 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     3361 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct
     3421 aaattctggc cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg
     3481 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     3541 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     3601 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     3661 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     3721 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     3781 atggagggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     3841 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     3901 gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     3961 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     4021 cgcacctggt gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag
     4081 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct
     4141 aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     4201 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     4261 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     4321 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     4381 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4441 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4501 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4561 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     4621 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4681 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     4741 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4801 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4861 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4921 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     4981 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     5041 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5101 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5161 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5221 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     5281 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     5341 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     5401 ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
     5461 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
     5521 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
     5581 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
     5641 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
     5701 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
     5761 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5821 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5881 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5941 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     6001 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     6061 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     6121 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     6181 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     6241 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     6301 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
     6361 atcctaccat ccactcgaca cacccgccag cggccgc
//
