LOCUS       sequence-299537-        6380 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 6380)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6380)
  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-m4258314
FEATURES             Location/Qualifiers
     source          1..6380
                     /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..289
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    342..463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(353..372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     407..426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      578..624
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      654..790
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    835..906
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1132..1720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1212..1231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2220..3429
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3266..3285
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3433..4032
                     /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(3433..3452)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3929..3949
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4059..4107
                     /note="synthetic polyadenylation signal"
     CDS             join(4373..4441,4442..5233)
                     /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(4591..4610)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5246..5382
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5534..5605
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5610..5656)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5674..5722
                     /note="synthetic polyadenylation signal"
     enhancer        5803..6106
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6108..6310
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6261..6281
                     /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 gcaccatcac catcaccatt aagctagctt
      301 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      361 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      421 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      481 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      541 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
      601 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
      661 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
      721 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
      781 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
      841 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
      901 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
      961 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1021 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1921 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     1981 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2041 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2101 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2161 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2221 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
     2281 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
     2341 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     2401 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     2461 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     2521 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     2581 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
     2641 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     2701 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     2761 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     2821 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
     2881 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
     2941 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     3001 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     3061 acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt aggtgtgcgg
     3121 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     3181 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     3241 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     3301 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     3361 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     3421 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3481 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3541 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3601 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3661 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3721 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac
     3781 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3841 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg
     3901 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3961 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4021 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa
     4081 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac
     4141 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4201 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4261 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4321 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4381 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4441 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4501 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4561 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4621 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4681 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4741 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     4801 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4861 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4921 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     4981 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5041 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5101 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5161 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5221 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5281 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5341 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa
     5401 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     5461 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     5521 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     5581 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     5641 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     5701 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
     5761 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     5821 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     5881 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     5941 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     6001 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6061 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
     6121 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     6181 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     6241 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     6301 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
     6361 acacacccgc cagcggccgc
//
