LOCUS       sequence-300701-        6855 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 6855)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6855)
  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-m4252964
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    58..106
                     /note="synthetic polyadenylation signal"
     enhancer        187..490
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        492..694
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     645..665
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1021..1041
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1042..1083
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1152..1273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1163..1182)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1217..1236
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1388..1434
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1464..1600
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1645..1716
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1942..2530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2022..2041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3030..4239
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4076..4095
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4243..5271
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5298..5346
                     /note="synthetic polyadenylation signal"
     CDS             join(5612..5680,5681..6472)
                     /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(5830..5849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6485..6621
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6773..6844
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(join(6849..6855,1..40))
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
       61 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
      121 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
      181 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      241 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      301 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      361 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      421 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      481 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      541 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      601 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      661 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
      721 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgccaccat gagcaaaatg
      781 tctgacgtca agtgtacatc cgtagttctg ttatcagtgc ttcaacagct gcgggtggag
      841 agttcgtcaa agctctgggc ccagtgcgtg cagctgcaca atgatatatt gctcgcgaaa
      901 gatactacgg aagctttcga gaagatggtc agcttgctat ctgttctact gagcatgcaa
      961 ggagcagtag acatcaacaa actttgcgaa gagatgttag acaaccgagc caccctgcaa
     1021 gaaaaccttt actttcagtc ggggaaacct atacccaatc cacttctcgg cttagattct
     1081 acctaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt accttcagct
     1141 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1201 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1261 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1321 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     1381 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     1441 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     1501 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     1561 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     1621 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     1681 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     1741 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     1801 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     1861 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1921 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1981 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2041 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2101 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2161 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2221 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2281 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2341 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2401 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2461 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2701 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2761 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     2821 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     2881 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     2941 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     3001 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     3061 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     3121 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     3181 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     3241 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     3301 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     3361 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     3421 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     3481 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     3541 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     3601 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     3661 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     3721 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     3781 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     3841 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     3901 gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     3961 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     4021 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     4081 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     4141 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     4201 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa aaagcctgaa
     4261 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     4321 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     4381 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     4441 cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga gttcagcgag
     4501 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     4561 accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat cgctgccgcc
     4621 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     4681 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     4741 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     4801 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     4861 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     4921 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     4981 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct ccgggcgtat
     5041 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     5101 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     5161 cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt agaagtactc
     5221 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata agctagtatg
     5281 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     5341 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     5401 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     5461 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     5521 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     5581 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     5641 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5701 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     5761 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     5821 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5881 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     5941 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6001 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     6061 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6121 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     6181 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     6241 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6301 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6361 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6421 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     6481 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6541 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     6601 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     6661 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6721 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     6781 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     6841 tctcaattca aaaaa
//
