LOCUS       sequence-300616-        6410 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 6410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6410)
  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-m4252784
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      229..817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     718..737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1043..1114
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1159..1295)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1325..1371)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1486..1607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1523..1542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1577..1596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1660..1701)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1702..1722)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(2049..2251)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2078..2098)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2253..2556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2637..2685
                     /note="synthetic polyadenylation signal"
     terminator      2703..2749
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2754..2825
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2977..3113)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3126..3917,3918..3986))
                     /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     3749..3768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4252..4300
                     /note="synthetic polyadenylation signal"
     CDS             complement(4327..4926)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4410..4430)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4907..4926
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4930..6139)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5074..5093)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
       61 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
      121 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
      181 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      241 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      301 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      361 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      421 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      481 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      541 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      601 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      661 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      721 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      781 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      841 acggttcctg gccttttgct ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg
      901 agaacgggtg caatagcaag gaccagatcg aacttgactg acgcaattct gagcgtagtc
      961 gaaacccaga caagtcgatc tcgccaagta gctgacttga ctggttaatt aagtttcgat
     1021 agcccaaggt aaccaatgag attagggagg gaagaaagtt tgatgaacta cttctgaccc
     1081 ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga tcgaaatcga gagatcgcct
     1141 gcaggatagc tgacgacctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
     1201 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
     1261 agcgtcgcat caggcatttt tttctgcgcc aatgccggta gatagcaatc cggcccgagg
     1321 ggcacaaaaa acccctcaag acccgtttag aggccccaag gggttatgct acgataccga
     1381 tagagatggg ccaataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1441 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1501 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1561 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtgag
     1621 ctgaaggtac gctgtatctc agtcagtcaa gctagcttag gtagaatcta agccgagaag
     1681 tggattgggt ataggtttcc ccgactgaaa gtaaaggttt tcttgcaggg tggctcggtt
     1741 gtctaacatc tcttcgcaaa gtttgttgat gtctactgct ccttgcatgc tcagtagaac
     1801 agatagcaag ctgaccatct tctcgaaagc ttccgtagta tctttcgcga gcaatatatc
     1861 attgtgcagc tgcacgcact gggcccagag ctttgacgaa ctctccaccc gcagctgttg
     1921 aagcactgat aacagaacta cggatgtaca cttgacgtca gacattttgc tcatggtggc
     1981 ggccgctggc gggtgtgtcg agtggatggt aggatcgaca aagatctgat ctgacggttc
     2041 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2101 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     2161 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     2221 cgcccattga tgtactgcca aaaccgcatc agcatggtaa tagcgatgac taatacgtag
     2281 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     2341 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     2401 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     2461 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     2521 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     2581 actaatgacc ccgtaattga ttactagatc tggctacact cgatcagcag ttagataata
     2641 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgcaacgatt
     2701 gatagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttga attgagaggt
     2761 agctgaagct gctgttatga ccactagagg gaagaagata cctgtggagc taatggtcca
     2821 agatgtcggg agccgcgatc gctgccaaaa aggtaagctg acgatagctg acgagattcg
     2881 atgcccgacg agcaaatgag cgccacggct gttcgcgtaa gctcgaaacg acggagattc
     2941 gatagagcca gagtcgagtt taaacgattc gggatcttaa ggataaattt ctggtaagga
     3001 ggacacgtat ggaagtgggc aagttgggga agccgtatcc gttgctgaat ctggcatatg
     3061 tgggagtata agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa tgcacctaga
     3121 atcggttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3181 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3241 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3301 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3361 atccagtcta tcaactgttg ccgggaagct agagtaagta gttcgccagt taatagttta
     3421 cgcaaggttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3481 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3541 aaagcggtta gctccttcgg tcctccaatc gttgtcagaa gtaagttggc cgcagtgtta
     3601 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3661 ttttctgtga ctggtgaata ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3721 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3781 gtgctcatca ttggaaagcg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3841 agatccagtt cgatgtaacc cactcgcgca cccaactgat cttcagcatc ttttactttc
     3901 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3961 gcgacacgga aatgttgaat actcatgcga aagcctcctg ctgatcctgt ctcttgatag
     4021 atgtttgcaa aagctaggac tcgattatac gttgaccgat cagactatca atatctaggt
     4081 ttaaacagat cggtacgctg ccgtgaccag gcttgagtac aatatctgac gattggactt
     4141 cctgggtccc aagctggatt ccgactggct gactgccaag caacgatatg gatcgtcgag
     4201 actcgcactg agaggatcgt tctcggtaca ccaggcgcgc ctagcgttac acacacaaaa
     4261 aaccaacaca catccatctt cgatggatag cgattttatt atctaagact aggcttacat
     4321 actagcctag gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc
     4381 gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga
     4441 ggtctccagg aaggcgggca ccccggcgcg ctcggcggcc tccactccgg ggagcacgac
     4501 ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca
     4561 cgcgggctcc ttgggccggt gcggcgccag gaggccctcc atctgttgct gcgcggccag
     4621 ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc
     4681 gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt
     4741 gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat
     4801 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
     4861 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
     4921 ggtcatgttg tctaacaaaa aagccaaaaa cggccagaat ttagcggaca atttacaagt
     4981 ctaacactga aaattacata ttgacccaaa tgattacatt tcaaaaggtg cctaaaaaac
     5041 ttcacaaaac acactcgcca accccgagcg catagttcaa aaccggagct tcagctactt
     5101 aagaagatag gtacataaaa ccgaccaaag aaactgacgc ctcacttatc cctcccctca
     5161 ccagaggtcc ggcgcctgtc gattcaggag agcctaccct aggcccgaac cctgcgtcct
     5221 gcgacggaga aaagcctacc gcacacctac cggctggtgg ccccaccctg cattataagc
     5281 caacagaacg ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg tacgggtgca
     5341 ctgcccaacg gcaccgccat aactgccgcc cccgcaacag acgacaaacc gagttctcca
     5401 gtcagtgaca aacttcacgt cagggtcccc agatggtgcc cagcccatct cacccgaata
     5461 agagctttcc cgcattagcg aagggctcaa gaccttgggt tcttgccgcc caccatgccc
     5521 cccaccttgt ttcaacgacc tcacagcccg cctcacaagc gtcttccatt caagactcgg
     5581 gaacagccgc cattttgctg cgctcccccc aacccccagt tcagggcaac cttgctcgcg
     5641 gacccagact acagcccttg gcggtctctc cacacgcttc cgtcccaccg agcggcccgg
     5701 cggccacgaa agccccggcc agcccagcag cccgctactc accaagtgac gatcacagcg
     5761 atccacaaac aagaaccgcg acccaaatcc cggctgcgac ggaactagct gtgccacacc
     5821 cggcgcgtcc ttatataatc atcggcgttc accgccccac ggagatccct ccgcagaatc
     5881 gccgagaagg gactactttt cctcgcctgt tccgctctct ggaaagaaaa ccagtgccct
     5941 agagtcaccc aagtcccgtc ctaaaatgtc cttctgctga tactggggtt ctaaggccga
     6001 gtcttatgag cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag gacgctcctg
     6061 cgcccttcgt ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc gggaggcgcc
     6121 aaaacccggc gcggaggcca ggcgcgcccc gatttcgata ttcctcggat ctcggataga
     6181 tcgggattga ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg atattcggga
     6241 ttggagtaat acctggcgat aggcgctgcc aagcaatagc caaggcgcaa ggccggccaa
     6301 gagacttgcg atagttcgtt cgagaggtag cctgcaagat ggatcggacg aaaggtttga
     6361 agattttccg acgagggcct tggagagttc ggaaatttaa atgcaactat
//
