LOCUS       sequence-300630-        6852 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 6852)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6852)
  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-m4252917
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    29..77
                     /note="synthetic polyadenylation signal"
     terminator      95..141
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    146..217
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(369..505)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(518..1309,1310..1378))
                     /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     1141..1160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1644..1692
                     /note="synthetic polyadenylation signal"
     CDS             complement(1719..2747)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2751..3960)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2895..2914)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4460..5048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4949..4968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5274..5345
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5390..5526)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5556..5602)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5717..5838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5754..5773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5808..5827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5907..5951)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(5952..5972)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6293..6495)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6322..6342)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6497..6800
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tctggctaca ctcgatcagc agttagataa taaaatcgct atccatcgaa gatggatgtg
       61 tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg ggcctctaaa
      121 cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat gaccactaga
      181 gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga tcgctgccaa
      241 aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg agcgccacgg
      301 ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag tttaaacgat
      361 tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg
      421 gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat
      481 caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta atcagtgagg
      541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
      661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt tgccgggaag
      781 ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt gctacaggca
      841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccaa
      961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa tactcaacca
     1081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag cgttcttcgg
     1201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgcg
     1261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatgc
     1381 gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg actcgattat
     1441 acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc tgccgtgacc
     1501 aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga ttccgactgg
     1561 ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc gttctcggta
     1621 caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
     1681 agcgatttta ttatctaaga ctaggcttac atactagctt attcctttgc cctcggacga
     1741 gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg
     1801 gcggcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt
     1861 gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga
     1921 tagagttggt caagaccaat gcggagcata tacgcccgga gccggggcga tcctgcaagc
     1981 tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc
     2041 cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca
     2101 atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc
     2161 acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc
     2221 gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca
     2281 gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat
     2341 gggccgaacc cgctcgtctg gctaagatcg gcggcagcga tggcatccat tgcctccgcg
     2401 accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt
     2461 gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc
     2521 ggaatcggga gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca
     2581 tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca
     2641 cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc
     2701 agaaacttct cgacagacgt cgcggtgagt tcaggctttt tacccatgtt gtctaacaaa
     2761 aaagccaaaa acggccagaa tttagcggac aatttacaag tctaacactg aaaattacat
     2821 attgacccaa atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa cacactcgcc
     2881 aaccccgagc gcatagttca aaaccggagc ttcagctact taagaagata ggtacataaa
     2941 accgaccaaa gaaactgacg cctcacttat ccctcccctc accagaggtc cggcgcctgt
     3001 cgattcagga gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag aaaagcctac
     3061 cgcacaccta ccggctggtg gccccaccct gcattataag ccaacagaac gggtgacgtc
     3121 acgacacgac gagggcgcgc gctcccaaag gtacgggtgc actgcccaac ggcaccgcca
     3181 taactgccgc ccccgcaaca gacgacaaac cgagttctcc agtcagtgac aaacttcacg
     3241 tcagggtccc cagatggtgc ccagcccatc tcacccgaat aagagctttc ccgcattagc
     3301 gaagggctca agaccttggg ttcttgccgc ccaccatgcc ccccaccttg tttcaacgac
     3361 ctcacagccc gcctcacaag cgtcttccat tcaagactcg ggaacagccg ccattttgct
     3421 gcgctccccc caacccccag ttcagggcaa ccttgctcgc ggacccagac tacagccctt
     3481 ggcggtctct ccacacgctt ccgtcccacc gagcggcccg gcggccacga aagccccggc
     3541 cagcccagca gcccgctact caccaagtga cgatcacagc gatccacaaa caagaaccgc
     3601 gacccaaatc ccggctgcga cggaactagc tgtgccacac ccggcgcgtc cttatataat
     3661 catcggcgtt caccgcccca cggagatccc tccgcagaat cgccgagaag ggactacttt
     3721 tcctcgcctg ttccgctctc tggaaagaaa accagtgccc tagagtcacc caagtcccgt
     3781 cctaaaatgt ccttctgctg atactggggt tctaaggccg agtcttatga gcagcgggcc
     3841 gctgtcctga gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg tctgacgtgg
     3901 cagcgctcgc cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg cgcggaggcc
     3961 aggcgcgccc cgatttcgat attcctcgga tctcggatag atcgggattg aggatgtagg
     4021 ttgaacttct ctcgggtgat cccgatctcg gatattcggg attggagtaa tacctggcga
     4081 taggcgctgc caagcaatag ccaaggcgca aggccggcca agagacttgc gatagttcgt
     4141 tcgagaggta gcctgcaaga tggatcggac gaaaggtttg aagattttcc gacgagggcc
     4201 ttggagagtt cggaaattta aatgcaacta tggatgaacg aaatagacag atcgctgaga
     4261 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4321 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4381 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4441 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4501 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4561 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4621 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4681 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4741 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4801 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4861 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4921 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4981 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5041 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5101 ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa ggaccagatc
     5161 gaacttgact gacgcaattc tgagcgtagt cgaaacccag acaagtcgat ctcgccaagt
     5221 agctgacttg actggttaat taagtttcga tagcccaagg taaccaatga gattagggag
     5281 ggaagaaagt ttgatgaact acttctgacc cctagtggtg tccagaaaag accattaaag
     5341 gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag ctgacgacct taaggataaa
     5401 tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg
     5461 aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc
     5521 caatgccggt agatagcaat ccggcccgag gggcacaaaa aacccctcaa gacccgttta
     5581 gaggccccaa ggggttatgc tacgataccg atagagatgg gccaatacca catttgtaga
     5641 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5701 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5761 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5821 actcatcaat gtatcttatc atgtctgtga gctgaaggta cgctgtatct cagtcagtca
     5881 agctagcaga ggcctgcgga tccttaactg tccatgtgct gcctctcgaa cttagctgcc
     5941 gcggtttctt tcgactgaaa gtaaaggttt tctaaggttg ccctgttgtc cagcatttct
     6001 tcacaaagct tgtttatgtc tacagcaccc tgcatggaaa gcaaaacaga aagtagtgaa
     6061 accatttttt caaaggcttc agtagtatct ttagctaaga gaatgtcatt gtgtaactgg
     6121 acacattgag cccacaattt agatgatgat tctactctga gttgttgcaa aactgagagt
     6181 aagactactg atgtgcactt tacatctgac attttcatgg tggcggccgc tggcgggtgt
     6241 gtcgagtgga tggtaggatc gacaaagatc tgatctgacg gttcactaaa ccagctctgc
     6301 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     6361 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     6421 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     6481 gccaaaaccg catcagcatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     6541 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     6601 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     6661 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     6721 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     6781 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     6841 ttgattacta ga
//
