LOCUS       sequence-300748-        6876 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 6876)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6876)
  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-m4252906
FEATURES             Location/Qualifiers
     source          1..6876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    26..74
                     /note="synthetic polyadenylation signal"
     terminator      92..138
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    143..214
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(366..502)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(515..1306,1307..1375))
                     /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     1138..1157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1641..1689
                     /note="synthetic polyadenylation signal"
     CDS             complement(1716..2744)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2748..3957)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2892..2911)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4457..5045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4946..4965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5271..5342
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5387..5523)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5553..5599)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5714..5835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5751..5770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5805..5824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5904..5969)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(5973..5993)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6314..6516)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6343..6363)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6518..6821
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat ggatgtgtgt
       61 tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc ctctaaacgg
      121 gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac cactagaggg
      181 aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg ctgccaaaaa
      241 ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc gccacggctg
      301 ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt aaacgattcg
      361 ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca agttggggaa
      421 gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc gtcgcatcag
      481 gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc agtgaggcac
      541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc cgggaagcta
      781 gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct acaggcatcg
      841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccaatcg
      961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac tcaaccaagt
     1081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt tcttcggggc
     1201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgcgcac
     1261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatgcgaa
     1381 agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact cgattatacg
     1441 ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc cgtgaccagg
     1501 cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc cgactggctg
     1561 actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt ctcggtacac
     1621 caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     1681 gattttatta tctaagacta ggcttacata ctagcttatt cctttgccct cggacgagtg
     1741 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcg
     1801 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg
     1861 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag
     1921 agttggtcaa gaccaatgcg gagcatatac gcccggagcc ggggcgatcc tgcaagctcc
     1981 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag
     2041 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg
     2101 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg
     2161 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg
     2221 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca
     2281 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg
     2341 ccgaacccgc tcgtctggct aagatcggcg gcagcgatgg catccattgc ctccgcgacc
     2401 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca
     2461 cggcgggaga tgcaataggt caggctctcg ctgaactccc caatgtcaag cacttccgga
     2521 atcgggagcg cggcggatgc aaagtgccga taaacataac gatctttgta gaaaccatcg
     2581 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga
     2641 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga
     2701 aacttctcga cagacgtcgc ggtgagttca ggctttttac ccatgttgtc taacaaaaaa
     2761 gccaaaaacg gccagaattt agcggacaat ttacaagtct aacactgaaa attacatatt
     2821 gacccaaatg attacatttc aaaaggtgcc taaaaaactt cacaaaacac actcgccaac
     2881 cccgagcgca tagttcaaaa ccggagcttc agctacttaa gaagataggt acataaaacc
     2941 gaccaaagaa actgacgcct cacttatccc tcccctcacc agaggtccgg cgcctgtcga
     3001 ttcaggagag cctaccctag gcccgaaccc tgcgtcctgc gacggagaaa agcctaccgc
     3061 acacctaccg gctggtggcc ccaccctgca ttataagcca acagaacggg tgacgtcacg
     3121 acacgacgag ggcgcgcgct cccaaaggta cgggtgcact gcccaacggc accgccataa
     3181 ctgccgcccc cgcaacagac gacaaaccga gttctccagt cagtgacaaa cttcacgtca
     3241 gggtccccag atggtgccca gcccatctca cccgaataag agctttcccg cattagcgaa
     3301 gggctcaaga ccttgggttc ttgccgccca ccatgccccc caccttgttt caacgacctc
     3361 acagcccgcc tcacaagcgt cttccattca agactcggga acagccgcca ttttgctgcg
     3421 ctccccccaa cccccagttc agggcaacct tgctcgcgga cccagactac agcccttggc
     3481 ggtctctcca cacgcttccg tcccaccgag cggcccggcg gccacgaaag ccccggccag
     3541 cccagcagcc cgctactcac caagtgacga tcacagcgat ccacaaacaa gaaccgcgac
     3601 ccaaatcccg gctgcgacgg aactagctgt gccacacccg gcgcgtcctt atataatcat
     3661 cggcgttcac cgccccacgg agatccctcc gcagaatcgc cgagaaggga ctacttttcc
     3721 tcgcctgttc cgctctctgg aaagaaaacc agtgccctag agtcacccaa gtcccgtcct
     3781 aaaatgtcct tctgctgata ctggggttct aaggccgagt cttatgagca gcgggccgct
     3841 gtcctgagcg tccgggcgga aggatcagga cgctcctgcg cccttcgtct gacgtggcag
     3901 cgctcgccgt gacgaggggg gcgcccgcgg gaggcgccaa aacccggcgc ggaggccagg
     3961 cgcgccccga tttcgatatt cctcggatct cggatagatc gggattgagg atgtaggttg
     4021 aacttctctc gggtgatccc gatctcggat attcgggatt ggagtaatac ctggcgatag
     4081 gcgctgccaa gcaatagcca aggcgcaagg ccggccaaga gacttgcgat agttcgttcg
     4141 agaggtagcc tgcaagatgg atcggacgaa aggtttgaag attttccgac gagggccttg
     4201 gagagttcgg aaatttaaat gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4261 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4321 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4381 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4441 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4501 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4561 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4621 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4681 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4741 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4801 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4861 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4921 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4981 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5041 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5101 acatgattta aatgaaactc gtaatcggag aacgggtgca atagcaagga ccagatcgaa
     5161 cttgactgac gcaattctga gcgtagtcga aacccagaca agtcgatctc gccaagtagc
     5221 tgacttgact ggttaattaa gtttcgatag cccaaggtaa ccaatgagat tagggaggga
     5281 agaaagtttg atgaactact tctgacccct agtggtgtcc agaaaagacc attaaaggaa
     5341 tgtcttgatc gaaatcgaga gatcgcctgc aggatagctg acgaccttaa ggataaattt
     5401 ctggtaagga ggacacgtat ggaagtgggc aagttgggga agccgtatcc gttgctgaat
     5461 ctggcatatg tgggagtata agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa
     5521 tgccggtaga tagcaatccg gcccgagggg cacaaaaaac ccctcaagac ccgtttagag
     5581 gccccaaggg gttatgctac gataccgata gagatgggcc aataccacat ttgtagaggt
     5641 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5701 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5761 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5821 catcaatgta tcttatcatg tctgtgagct gaaggtacgc tgtatctcag tcagtcaagc
     5881 tagcagaggc ctgcggatcc ttatttgtca tcgtcgtcct tgtaatcaat gtcatgatct
     5941 ttgtagtctc catcgtggtc cttataatcc atcgactgaa agtaaaggtt ttctaaggtt
     6001 gccctgttgt ccagcatttc ttcacaaagc ttgtttatgt ctacagcacc ctgcatggaa
     6061 agcaaaacag aaagtagtga aaccattttt tcaaaggctt cagtagtatc tttagctaag
     6121 agaatgtcat tgtgtaactg gacacattga gcccacaatt tagatgatga ttctactctg
     6181 agttgttgca aaactgagag taagactact gatgtgcact ttacatctga cattttcatg
     6241 gtggcggccg ctggcgggtg tgtcgagtgg atggtaggat cgacaaagat ctgatctgac
     6301 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     6361 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     6421 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     6481 atccacgccc attgatgtac tgccaaaacc gcatcagcat ggtaatagcg atgactaata
     6541 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     6601 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     6661 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     6721 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     6781 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     6841 tatgaactaa tgaccccgta attgattact agatct
//
