LOCUS       sequence-297966-        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-m4134097
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             915..935
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             936..962
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1031..1152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1042..1061)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1096..1115
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1267..1313
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1343..1479
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1524..1595
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1821..2409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1901..1920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2909..4118
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3955..3974
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4122..5150
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5177..5225
                     /note="synthetic polyadenylation signal"
     CDS             join(5491..5559,5560..6351)
                     /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(5709..5728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6364..6500
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6652..6723
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6728..6774)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6792..6840
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca
      661 ccatgagcaa aatgtctgac gtcaagtgta catccgtagt tctgttatca gtgcttcaac
      721 agctgcgggt ggagagttcg tcaaagctct gggcccagtg cgtgcagctg cacaatgata
      781 tattgctcgc gaaagatact acggaagctt tcgagaagat ggtcagcttg ctatctgttc
      841 tactgagcat gcaaggagca gtagacatca acaaactttg cgaagagatg ttagacaacc
      901 gagccaccct gcaagaaaac ctttactttc agtcgtaccc ctatgacgta ccagattatg
      961 cgtaaggatc cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc
     1021 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1081 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1141 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1201 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
     1261 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
     1321 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     1381 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     1441 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
     1501 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     1561 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     1621 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
     1681 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
     1741 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2641 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     2701 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     2761 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     2821 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     2881 cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg
     2941 ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg
     3001 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     3061 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     3121 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     3181 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     3241 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     3301 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     3361 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     3421 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     3481 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     3541 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga
     3601 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     3661 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     3721 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     3781 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     3841 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     3901 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     3961 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     4021 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt
     4081 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac
     4141 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     4201 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     4261 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     4321 actttgcatc cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga
     4381 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     4441 ccgaactgcc cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg
     4501 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     4561 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     4621 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     4681 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     4741 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4801 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     4861 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata
     4921 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4981 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     5041 gtacacaaat cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg
     5101 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt
     5161 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     5221 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     5281 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     5341 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     5401 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     5461 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     5521 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5581 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5641 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     5701 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5761 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     5821 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5881 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     5941 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6001 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     6061 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     6121 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6181 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6241 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6301 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     6361 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     6421 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     6481 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     6541 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6601 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6661 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     6721 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     6781 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
//
