LOCUS       sequence-300437-        6347 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 6347)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6347)
  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-m4252978
FEATURES             Location/Qualifiers
     source          1..6347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      52..98
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    103..174
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(326..462)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(475..1266,1267..1335))
                     /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     1098..1117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1601..1649
                     /note="synthetic polyadenylation signal"
     CDS             complement(1676..2275)
                     /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(1759..1779)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2256..2275
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2279..3488)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2423..2442)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      3988..4576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4477..4496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4802..4873
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4918..5054)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5084..5130)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5245..5366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5282..5301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5336..5355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(5745..5947)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5774..5794)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5949..6252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6333..6347,1..34)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gcaacgattg atagcataac
       61 cccttggggc ctctaaacgg gtcttgaggg gttttttgaa ttgagaggta gctgaagctg
      121 ctgttatgac cactagaggg aagaagatac ctgtggagct aatggtccaa gatgtcggga
      181 gccgcgatcg ctgccaaaaa ggtaagctga cgatagctga cgagattcga tgcccgacga
      241 gcaaatgagc gccacggctg ttcgcgtaag ctcgaaacga cggagattcg atagagccag
      301 agtcgagttt aaacgattcg ggatcttaag gataaatttc tggtaaggag gacacgtatg
      361 gaagtgggca agttggggaa gccgtatccg ttgctgaatc tggcatatgt gggagtataa
      421 gacgcgcagc gtcgcatcag gcattttttt ctgcgccaat gcacctagaa tcggttacca
      481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      721 caactgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttac gcaaggttgt
      781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      901 ctccttcggt cctccaatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1021 tggtgaatac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1141 tggaaagcgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1201 gatgtaaccc actcgcgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1321 atgttgaata ctcatgcgaa agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa
     1381 agctaggact cgattatacg ttgaccgatc agactatcaa tatctaggtt taaacagatc
     1441 ggtacgctgc cgtgaccagg cttgagtaca atatctgacg attggacttc ctgggtccca
     1501 agctggattc cgactggctg actgccaagc aacgatatgg atcgtcgaga ctcgcactga
     1561 gaggatcgtt ctcggtacac caggcgcgcc tagcgttaca cacacaaaaa accaacacac
     1621 atccatcttc gatggatagc gattttatta tctaagacta ggcttacata ctagcctagg
     1681 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg
     1741 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga
     1801 aggcgggcac cccggcgcgc tcggcggcct ccactccggg gagcacgacg gcgctgccca
     1861 gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct
     1921 tgggccggtg cggcgccagg aggccctcca tctgttgctg cgcggccagc cgggaaccgc
     1981 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg
     2041 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga
     2101 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg
     2161 gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
     2221 ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatgttgt
     2281 ctaacaaaaa agccaaaaac ggccagaatt tagcggacaa tttacaagtc taacactgaa
     2341 aattacatat tgacccaaat gattacattt caaaaggtgc ctaaaaaact tcacaaaaca
     2401 cactcgccaa ccccgagcgc atagttcaaa accggagctt cagctactta agaagatagg
     2461 tacataaaac cgaccaaaga aactgacgcc tcacttatcc ctcccctcac cagaggtccg
     2521 gcgcctgtcg attcaggaga gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa
     2581 aagcctaccg cacacctacc ggctggtggc cccaccctgc attataagcc aacagaacgg
     2641 gtgacgtcac gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg
     2701 caccgccata actgccgccc ccgcaacaga cgacaaaccg agttctccag tcagtgacaa
     2761 acttcacgtc agggtcccca gatggtgccc agcccatctc acccgaataa gagctttccc
     2821 gcattagcga agggctcaag accttgggtt cttgccgccc accatgcccc ccaccttgtt
     2881 tcaacgacct cacagcccgc ctcacaagcg tcttccattc aagactcggg aacagccgcc
     2941 attttgctgc gctcccccca acccccagtt cagggcaacc ttgctcgcgg acccagacta
     3001 cagcccttgg cggtctctcc acacgcttcc gtcccaccga gcggcccggc ggccacgaaa
     3061 gccccggcca gcccagcagc ccgctactca ccaagtgacg atcacagcga tccacaaaca
     3121 agaaccgcga cccaaatccc ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct
     3181 tatataatca tcggcgttca ccgccccacg gagatccctc cgcagaatcg ccgagaaggg
     3241 actacttttc ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta gagtcaccca
     3301 agtcccgtcc taaaatgtcc ttctgctgat actggggttc taaggccgag tcttatgagc
     3361 agcgggccgc tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc
     3421 tgacgtggca gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg
     3481 cggaggccag gcgcgccccg atttcgatat tcctcggatc tcggatagat cgggattgag
     3541 gatgtaggtt gaacttctct cgggtgatcc cgatctcgga tattcgggat tggagtaata
     3601 cctggcgata ggcgctgcca agcaatagcc aaggcgcaag gccggccaag agacttgcga
     3661 tagttcgttc gagaggtagc ctgcaagatg gatcggacga aaggtttgaa gattttccga
     3721 cgagggcctt ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa atagacagat
     3781 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3841 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3901 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3961 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4021 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4081 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4141 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4201 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4261 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4321 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4381 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4441 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4501 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4561 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4621 gccttttgct cacatgattt aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg
     4681 accagatcga acttgactga cgcaattctg agcgtagtcg aaacccagac aagtcgatct
     4741 cgccaagtag ctgacttgac tggttaatta agtttcgata gcccaaggta accaatgaga
     4801 ttagggaggg aagaaagttt gatgaactac ttctgacccc tagtggtgtc cagaaaagac
     4861 cattaaagga atgtcttgat cgaaatcgag agatcgcctg caggatagct gacgacctta
     4921 aggataaatt tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc
     4981 cgttgctgaa tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt
     5041 ttctgcgcca atgccggtag atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga
     5101 cccgtttaga ggccccaagg ggttatgcta cgataccgat agagatgggc caataccaca
     5161 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5221 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5341 ttgtccaaac tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca
     5401 gtcagtcaag ctagcttatt gcagggtggc tcggttgtct aacatctctt cgcaaagttt
     5461 gttgatgtct actgctcctt gcatgctcag tagaacagat agcaagctga ccatcttctc
     5521 gaaagcttcc gtagtatctt tcgcgagcaa tatatcattg tgcagctgca cgcactgggc
     5581 ccagagcttt gacgaactct ccacccgcag ctgttgaagc actgataaca gaactacgga
     5641 tgtacacttg acgtcagaca ttttgctcat ggtggcggcc gctggcgggt gtgtcgagtg
     5701 gatggtagga tcgacaaaga tctgatctga cggttcacta aaccagctct gcttatatag
     5761 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     5821 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     5881 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     5941 cgcatcagca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6001 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6061 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     6121 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     6181 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     6241 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     6301 tagatctggc tacactcgat cagcagttag ataataaaat cgctatc
//
