LOCUS       sequence-300604-        6858 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 6858)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6858)
  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-m4252881
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      221..809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     710..729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1035..1106
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1151..1287)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1317..1363)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1478..1599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1515..1534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1569..1588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1668..1712)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1713..1733)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(2060..2262)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2089..2109)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2264..2567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2648..2696
                     /note="synthetic polyadenylation signal"
     terminator      2714..2760
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2765..2836
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2988..3124)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3137..3928,3929..3997))
                     /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     3760..3779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4263..4311
                     /note="synthetic polyadenylation signal"
     CDS             complement(4338..5366)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5370..6579)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5514..5533)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
       61 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
      121 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
      181 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
      241 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
      301 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
      361 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
      421 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
      481 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      541 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      601 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      661 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      721 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      781 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      841 tggccttttg ctggcctttt gctcacatga tttaaatgaa actcgtaatc ggagaacggg
      901 tgcaatagca aggaccagat cgaacttgac tgacgcaatt ctgagcgtag tcgaaaccca
      961 gacaagtcga tctcgccaag tagctgactt gactggttaa ttaagtttcg atagcccaag
     1021 gtaaccaatg agattaggga gggaagaaag tttgatgaac tacttctgac ccctagtggt
     1081 gtccagaaaa gaccattaaa ggaatgtctt gatcgaaatc gagagatcgc ctgcaggata
     1141 gctgacgacc ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg
     1201 gggaagccgt atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc
     1261 atcaggcatt tttttctgcg ccaatgccgg tagatagcaa tccggcccga ggggcacaaa
     1321 aaacccctca agacccgttt agaggcccca aggggttatg ctacgatacc gatagagatg
     1381 ggccaatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1441 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgtg agctgaaggt
     1621 acgctgtatc tcagtcagtc aagctagcag aggcctgcgg atccttaact gtccatgtgc
     1681 tgcctctcga acttagctgc cgcggtttct ttcgactgaa agtaaaggtt ttcttgcagg
     1741 gtggctcggt tgtctaacat ctcttcgcaa agtttgttga tgtctactgc tccttgcatg
     1801 ctcagtagaa cagatagcaa gctgaccatc ttctcgaaag cttccgtagt atctttcgcg
     1861 agcaatatat cattgtgcag ctgcacgcac tgggcccaga gctttgacga actctccacc
     1921 cgcagctgtt gaagcactga taacagaact acggatgtac acttgacgtc agacattttg
     1981 ctcatggtgg cggccgctgg cgggtgtgtc gagtggatgg taggatcgac aaagatctga
     2041 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     2101 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     2161 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     2221 accgctatcc acgcccattg atgtactgcc aaaaccgcat cagcatggta atagcgatga
     2281 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     2341 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     2401 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     2461 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     2521 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     2581 atgggctatg aactaatgac cccgtaattg attactagat ctggctacac tcgatcagca
     2641 gttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     2701 acgcaacgat tgatagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2761 aattgagagg tagctgaagc tgctgttatg accactagag ggaagaagat acctgtggag
     2821 ctaatggtcc aagatgtcgg gagccgcgat cgctgccaaa aaggtaagct gacgatagct
     2881 gacgagattc gatgcccgac gagcaaatga gcgccacggc tgttcgcgta agctcgaaac
     2941 gacggagatt cgatagagcc agagtcgagt ttaaacgatt cgggatctta aggataaatt
     3001 tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa
     3061 tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca
     3121 atgcacctag aatcggttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3361 tatccgcctc catccagtct atcaactgtt gccgggaagc tagagtaagt agttcgccag
     3421 ttaatagttt acgcaaggtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccaat cgttgtcaga agtaagttgg
     3601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3661 ccgtaagatg cttttctgtg actggtgaat actcaaccaa gtcattctga gaatagtgta
     3721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3781 gaactttaaa agtgctcatc attggaaagc gttcttcggg gcgaaaactc tcaaggatct
     3841 taccgctgtt gagatccagt tcgatgtaac ccactcgcgc acccaactga tcttcagcat
     3901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3961 agggaataag ggcgacacgg aaatgttgaa tactcatgcg aaagcctcct gctgatcctg
     4021 tctcttgata gatgtttgca aaagctagga ctcgattata cgttgaccga tcagactatc
     4081 aatatctagg tttaaacaga tcggtacgct gccgtgacca ggcttgagta caatatctga
     4141 cgattggact tcctgggtcc caagctggat tccgactggc tgactgccaa gcaacgatat
     4201 ggatcgtcga gactcgcact gagaggatcg ttctcggtac accaggcgcg cctagcgtta
     4261 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaagac
     4321 taggcttaca tactagctta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca
     4381 ctatcggcga gtacttctac acagccatcg gtccagacgg cggcgcttct gcgggcgatt
     4441 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc
     4501 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg
     4561 cggagcatat acgcccggag ccggggcgat cctgcaagct ccggatgcct ccgctcgaag
     4621 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc
     4681 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca
     4741 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg
     4801 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg
     4861 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca
     4921 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg
     4981 ctaagatcgg cggcagcgat ggcatccatt gcctccgcga ccggctgcag aacagcgggc
     5041 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag
     5101 gtcaggctct cgctgaactc cccaatgtca agcacttccg gaatcgggag cgcggcggat
     5161 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
     5221 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
     5281 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
     5341 gcggtgagtt caggcttttt acccatgttg tctaacaaaa aagccaaaaa cggccagaat
     5401 ttagcggaca atttacaagt ctaacactga aaattacata ttgacccaaa tgattacatt
     5461 tcaaaaggtg cctaaaaaac ttcacaaaac acactcgcca accccgagcg catagttcaa
     5521 aaccggagct tcagctactt aagaagatag gtacataaaa ccgaccaaag aaactgacgc
     5581 ctcacttatc cctcccctca ccagaggtcc ggcgcctgtc gattcaggag agcctaccct
     5641 aggcccgaac cctgcgtcct gcgacggaga aaagcctacc gcacacctac cggctggtgg
     5701 ccccaccctg cattataagc caacagaacg ggtgacgtca cgacacgacg agggcgcgcg
     5761 ctcccaaagg tacgggtgca ctgcccaacg gcaccgccat aactgccgcc cccgcaacag
     5821 acgacaaacc gagttctcca gtcagtgaca aacttcacgt cagggtcccc agatggtgcc
     5881 cagcccatct cacccgaata agagctttcc cgcattagcg aagggctcaa gaccttgggt
     5941 tcttgccgcc caccatgccc cccaccttgt ttcaacgacc tcacagcccg cctcacaagc
     6001 gtcttccatt caagactcgg gaacagccgc cattttgctg cgctcccccc aacccccagt
     6061 tcagggcaac cttgctcgcg gacccagact acagcccttg gcggtctctc cacacgcttc
     6121 cgtcccaccg agcggcccgg cggccacgaa agccccggcc agcccagcag cccgctactc
     6181 accaagtgac gatcacagcg atccacaaac aagaaccgcg acccaaatcc cggctgcgac
     6241 ggaactagct gtgccacacc cggcgcgtcc ttatataatc atcggcgttc accgccccac
     6301 ggagatccct ccgcagaatc gccgagaagg gactactttt cctcgcctgt tccgctctct
     6361 ggaaagaaaa ccagtgccct agagtcaccc aagtcccgtc ctaaaatgtc cttctgctga
     6421 tactggggtt ctaaggccga gtcttatgag cagcgggccg ctgtcctgag cgtccgggcg
     6481 gaaggatcag gacgctcctg cgcccttcgt ctgacgtggc agcgctcgcc gtgacgaggg
     6541 gggcgcccgc gggaggcgcc aaaacccggc gcggaggcca ggcgcgcccc gatttcgata
     6601 ttcctcggat ctcggataga tcgggattga ggatgtaggt tgaacttctc tcgggtgatc
     6661 ccgatctcgg atattcggga ttggagtaat acctggcgat aggcgctgcc aagcaatagc
     6721 caaggcgcaa ggccggccaa gagacttgcg atagttcgtt cgagaggtag cctgcaagat
     6781 ggatcggacg aaaggtttga agattttccg acgagggcct tggagagttc ggaaatttaa
     6841 atgcaactat ggatgaac
//
