LOCUS       sequence-300749-        6389 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 6389)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6389)
  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-m4252824
FEATURES             Location/Qualifiers
     source          1..6389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    63..184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(100..119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     154..173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(237..263)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(264..284)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(605..807)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(634..654)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        809..1112
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1193..1241
                     /note="synthetic polyadenylation signal"
     terminator      1259..1305
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1310..1381
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1533..1669)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1682..2473,2474..2542))
                     /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     2305..2324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2808..2856
                     /note="synthetic polyadenylation signal"
     CDS             complement(2883..3482)
                     /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(2966..2986)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3463..3482
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3486..4695)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3630..3649)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5195..5783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5684..5703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6009..6080
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6125..6261)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6291..6337)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
       61 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      121 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      181 cttatcatgt ctgtgagctg aaggtacgct gtatctcagt cagtcaagct agcttacgca
      241 taatctggta cgtcataggg gtacgactga aagtaaaggt tttctaaggt tgccctgttg
      301 tccagcattt cttcacaaag cttgtttatg tctacagcac cctgcatgga aagcaaaaca
      361 gaaagtagtg aaaccatttt ttcaaaggct tcagtagtat ctttagctaa gagaatgtca
      421 ttgtgtaact ggacacattg agcccacaat ttagatgatg attctactct gagttgttgc
      481 aaaactgaga gtaagactac tgatgtgcac tttacatctg acattttcat ggtggcggcc
      541 gctggcgggt gtgtcgagtg gatggtagga tcgacaaaga tctgatctga cggttcacta
      601 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
      661 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
      721 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
      781 cattgatgta ctgccaaaac cgcatcagca tggtaatagc gatgactaat acgtagatgt
      841 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
      901 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
      961 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     1021 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     1081 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     1141 atgaccccgt aattgattac tagatctggc tacactcgat cagcagttag ataataaaat
     1201 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgca acgattgata
     1261 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgaattg agaggtagct
     1321 gaagctgctg ttatgaccac tagagggaag aagatacctg tggagctaat ggtccaagat
     1381 gtcgggagcc gcgatcgctg ccaaaaaggt aagctgacga tagctgacga gattcgatgc
     1441 ccgacgagca aatgagcgcc acggctgttc gcgtaagctc gaaacgacgg agattcgata
     1501 gagccagagt cgagtttaaa cgattcggga tcttaaggat aaatttctgg taaggaggac
     1561 acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg
     1621 agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgca cctagaatcg
     1681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1921 agtctatcaa ctgttgccgg gaagctagag taagtagttc gccagttaat agtttacgca
     1981 aggttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2101 cggttagctc cttcggtcct ccaatcgttg tcagaagtaa gttggccgca gtgttatcac
     2161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2221 ctgtgactgg tgaatactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2341 tcatcattgg aaagcgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2401 ccagttcgat gtaacccact cgcgcaccca actgatcttc agcatctttt actttcacca
     2461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2521 cacggaaatg ttgaatactc atgcgaaagc ctcctgctga tcctgtctct tgatagatgt
     2581 ttgcaaaagc taggactcga ttatacgttg accgatcaga ctatcaatat ctaggtttaa
     2641 acagatcggt acgctgccgt gaccaggctt gagtacaata tctgacgatt ggacttcctg
     2701 ggtcccaagc tggattccga ctggctgact gccaagcaac gatatggatc gtcgagactc
     2761 gcactgagag gatcgttctc ggtacaccag gcgcgcctag cgttacacac acaaaaaacc
     2821 aacacacatc catcttcgat ggatagcgat tttattatct aagactaggc ttacatacta
     2881 gcctaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     2941 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
     3001 tccaggaagg cgggcacccc ggcgcgctcg gcggcctcca ctccggggag cacgacggcg
     3061 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     3121 ggctccttgg gccggtgcgg cgccaggagg ccctccatct gttgctgcgc ggccagccgg
     3181 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     3241 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     3301 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     3361 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     3421 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     3481 atgttgtcta acaaaaaagc caaaaacggc cagaatttag cggacaattt acaagtctaa
     3541 cactgaaaat tacatattga cccaaatgat tacatttcaa aaggtgccta aaaaacttca
     3601 caaaacacac tcgccaaccc cgagcgcata gttcaaaacc ggagcttcag ctacttaaga
     3661 agataggtac ataaaaccga ccaaagaaac tgacgcctca cttatccctc ccctcaccag
     3721 aggtccggcg cctgtcgatt caggagagcc taccctaggc ccgaaccctg cgtcctgcga
     3781 cggagaaaag cctaccgcac acctaccggc tggtggcccc accctgcatt ataagccaac
     3841 agaacgggtg acgtcacgac acgacgaggg cgcgcgctcc caaaggtacg ggtgcactgc
     3901 ccaacggcac cgccataact gccgcccccg caacagacga caaaccgagt tctccagtca
     3961 gtgacaaact tcacgtcagg gtccccagat ggtgcccagc ccatctcacc cgaataagag
     4021 ctttcccgca ttagcgaagg gctcaagacc ttgggttctt gccgcccacc atgcccccca
     4081 ccttgtttca acgacctcac agcccgcctc acaagcgtct tccattcaag actcgggaac
     4141 agccgccatt ttgctgcgct ccccccaacc cccagttcag ggcaaccttg ctcgcggacc
     4201 cagactacag cccttggcgg tctctccaca cgcttccgtc ccaccgagcg gcccggcggc
     4261 cacgaaagcc ccggccagcc cagcagcccg ctactcacca agtgacgatc acagcgatcc
     4321 acaaacaaga accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc cacacccggc
     4381 gcgtccttat ataatcatcg gcgttcaccg ccccacggag atccctccgc agaatcgccg
     4441 agaagggact acttttcctc gcctgttccg ctctctggaa agaaaaccag tgccctagag
     4501 tcacccaagt cccgtcctaa aatgtccttc tgctgatact ggggttctaa ggccgagtct
     4561 tatgagcagc gggccgctgt cctgagcgtc cgggcggaag gatcaggacg ctcctgcgcc
     4621 cttcgtctga cgtggcagcg ctcgccgtga cgaggggggc gcccgcggga ggcgccaaaa
     4681 cccggcgcgg aggccaggcg cgccccgatt tcgatattcc tcggatctcg gatagatcgg
     4741 gattgaggat gtaggttgaa cttctctcgg gtgatcccga tctcggatat tcgggattgg
     4801 agtaatacct ggcgataggc gctgccaagc aatagccaag gcgcaaggcc ggccaagaga
     4861 cttgcgatag ttcgttcgag aggtagcctg caagatggat cggacgaaag gtttgaagat
     4921 tttccgacga gggccttgga gagttcggaa atttaaatgc aactatggat gaacgaaata
     4981 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5821 tttgctggcc ttttgctcac atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat
     5881 agcaaggacc agatcgaact tgactgacgc aattctgagc gtagtcgaaa cccagacaag
     5941 tcgatctcgc caagtagctg acttgactgg ttaattaagt ttcgatagcc caaggtaacc
     6001 aatgagatta gggagggaag aaagtttgat gaactacttc tgacccctag tggtgtccag
     6061 aaaagaccat taaaggaatg tcttgatcga aatcgagaga tcgcctgcag gatagctgac
     6121 gaccttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag
     6181 ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg
     6241 catttttttc tgcgccaatg ccggtagata gcaatccggc ccgaggggca caaaaaaccc
     6301 ctcaagaccc gtttagaggc cccaaggggt tatgctacga taccgataga gatgggccaa
     6361 taccacattt gtagaggttt tacttgctt
//
