LOCUS       sequence-297451-        6840 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 6840)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6840)
  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-m4252895
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             278..304
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    373..494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     438..457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      609..655
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      685..821
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    866..937
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1163..1751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1243..1262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2251..3460
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3297..3316
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3464..4492
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4519..4567
                     /note="synthetic polyadenylation signal"
     CDS             join(4833..4901,4902..5693)
                     /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(5051..5070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5706..5842
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5994..6065
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6070..6116)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6134..6182
                     /note="synthetic polyadenylation signal"
     enhancer        6263..6566
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6568..6770
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6721..6741
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaagaaa acctttactt tcagtcgtac ccctatgacg taccagatta
      301 tgcgtaagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
      361 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      421 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      481 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      541 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
      601 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
      661 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
      721 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
      781 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
      841 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
      901 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
      961 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     1021 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     1081 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1561 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1801 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1861 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1921 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1981 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     2041 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     2101 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     2161 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     2221 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     2281 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     2341 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     2401 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     2461 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     2521 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     2581 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     2641 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     2701 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     2761 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     2821 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     2881 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     2941 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     3001 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3061 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3121 agggtggggc caccagccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3181 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     3241 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     3301 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     3361 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     3421 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     3481 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     3541 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     3601 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     3661 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     3721 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     3781 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     3841 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     3901 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     3961 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4021 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4081 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4141 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4201 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     4261 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     4321 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     4381 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     4441 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     4501 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     4561 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     4621 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     4681 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     4741 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     4801 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     4861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4921 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     4981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     5041 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5101 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     5161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5221 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     5281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     5401 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     5461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5521 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     5701 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     5761 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     5821 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     5881 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     5941 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     6001 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     6061 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     6121 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     6181 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     6241 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6301 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6361 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6421 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6481 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6541 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     6601 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6661 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6721 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     6781 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
