LOCUS       sequence-300496-        6341 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 6341)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6341)
  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-m4252970
FEATURES             Location/Qualifiers
     source          1..6341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    74..122
                     /note="synthetic polyadenylation signal"
     CDS             complement(149..748)
                     /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(232..252)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     729..748
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(752..1961)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(896..915)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      2461..3049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2950..2969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3275..3346
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3391..3527)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3557..3603)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3718..3839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3755..3774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3809..3828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(4212..4414)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4241..4261)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4416..4719
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4800..4848
                     /note="synthetic polyadenylation signal"
     terminator      4866..4912
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    4917..4988
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5140..5276)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(5289..6080,6081..6149))
                     /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     5912..5931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agcaacgata tggatcgtcg agactcgcac tgagaggatc gttctcggta caccaggcgc
       61 gcctagcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
      121 ttatctaaga ctaggcttac atactagcct aggcaccggg cttgcgggtc atgcaccagg
      181 tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga
      241 aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggcgg
      301 cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga
      361 cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggccct
      421 ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct
      481 cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc
      541 cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt
      601 gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt
      661 agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg cgggtggcga
      721 ggcgcaccgt gggcttgtac tcggtcatgt tgtctaacaa aaaagccaaa aacggccaga
      781 atttagcgga caatttacaa gtctaacact gaaaattaca tattgaccca aatgattaca
      841 tttcaaaagg tgcctaaaaa acttcacaaa acacactcgc caaccccgag cgcatagttc
      901 aaaaccggag cttcagctac ttaagaagat aggtacataa aaccgaccaa agaaactgac
      961 gcctcactta tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc
     1021 ctaggcccga accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggctggt
     1081 ggccccaccc tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg
     1141 cgctcccaaa ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac
     1201 agacgacaaa ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg
     1261 cccagcccat ctcacccgaa taagagcttt cccgcattag cgaagggctc aagaccttgg
     1321 gttcttgccg cccaccatgc cccccacctt gtttcaacga cctcacagcc cgcctcacaa
     1381 gcgtcttcca ttcaagactc gggaacagcc gccattttgc tgcgctcccc ccaaccccca
     1441 gttcagggca accttgctcg cggacccaga ctacagccct tggcggtctc tccacacgct
     1501 tccgtcccac cgagcggccc ggcggccacg aaagccccgg ccagcccagc agcccgctac
     1561 tcaccaagtg acgatcacag cgatccacaa acaagaaccg cgacccaaat cccggctgcg
     1621 acggaactag ctgtgccaca cccggcgcgt ccttatataa tcatcggcgt tcaccgcccc
     1681 acggagatcc ctccgcagaa tcgccgagaa gggactactt ttcctcgcct gttccgctct
     1741 ctggaaagaa aaccagtgcc ctagagtcac ccaagtcccg tcctaaaatg tccttctgct
     1801 gatactgggg ttctaaggcc gagtcttatg agcagcgggc cgctgtcctg agcgtccggg
     1861 cggaaggatc aggacgctcc tgcgcccttc gtctgacgtg gcagcgctcg ccgtgacgag
     1921 gggggcgccc gcgggaggcg ccaaaacccg gcgcggaggc caggcgcgcc ccgatttcga
     1981 tattcctcgg atctcggata gatcgggatt gaggatgtag gttgaacttc tctcgggtga
     2041 tcccgatctc ggatattcgg gattggagta atacctggcg ataggcgctg ccaagcaata
     2101 gccaaggcgc aaggccggcc aagagacttg cgatagttcg ttcgagaggt agcctgcaag
     2161 atggatcgga cgaaaggttt gaagattttc cgacgagggc cttggagagt tcggaaattt
     2221 aaatgcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2281 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2341 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2401 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2461 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2521 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2581 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2641 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2701 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2761 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2821 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2881 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2941 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3001 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3061 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatga tttaaatgaa
     3121 actcgtaatc ggagaacggg tgcaatagca aggaccagat cgaacttgac tgacgcaatt
     3181 ctgagcgtag tcgaaaccca gacaagtcga tctcgccaag tagctgactt gactggttaa
     3241 ttaagtttcg atagcccaag gtaaccaatg agattaggga gggaagaaag tttgatgaac
     3301 tacttctgac ccctagtggt gtccagaaaa gaccattaaa ggaatgtctt gatcgaaatc
     3361 gagagatcgc ctgcaggata gctgacgacc ttaaggataa atttctggta aggaggacac
     3421 gtatggaagt gggcaagttg gggaagccgt atccgttgct gaatctggca tatgtgggag
     3481 tataagacgc gcagcgtcgc atcaggcatt tttttctgcg ccaatgccgg tagatagcaa
     3541 tccggcccga ggggcacaaa aaacccctca agacccgttt agaggcccca aggggttatg
     3601 ctacgatacc gatagagatg ggccaatacc acatttgtag aggttttact tgctttaaaa
     3661 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3841 catgtctgtg agctgaaggt acgctgtatc tcagtcagtc aagctagctt ataaggttgc
     3901 cctgttgtcc agcatttctt cacaaagctt gtttatgtct acagcaccct gcatggaaag
     3961 caaaacagaa agtagtgaaa ccattttttc aaaggcttca gtagtatctt tagctaagag
     4021 aatgtcattg tgtaactgga cacattgagc ccacaattta gatgatgatt ctactctgag
     4081 ttgttgcaaa actgagagta agactactga tgtgcacttt acatctgaca ttttcatggt
     4141 ggcggccgct ggcgggtgtg tcgagtggat ggtaggatcg acaaagatct gatctgacgg
     4201 ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     4261 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     4321 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     4381 ccacgcccat tgatgtactg ccaaaaccgc atcagcatgg taatagcgat gactaatacg
     4441 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     4501 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     4561 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     4621 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     4681 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     4741 tgaactaatg accccgtaat tgattactag atctggctac actcgatcag cagttagata
     4801 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgcaacg
     4861 attgatagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaattgaga
     4921 ggtagctgaa gctgctgtta tgaccactag agggaagaag atacctgtgg agctaatggt
     4981 ccaagatgtc gggagccgcg atcgctgcca aaaaggtaag ctgacgatag ctgacgagat
     5041 tcgatgcccg acgagcaaat gagcgccacg gctgttcgcg taagctcgaa acgacggaga
     5101 ttcgatagag ccagagtcga gtttaaacga ttcgggatct taaggataaa tttctggtaa
     5161 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
     5221 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgcacct
     5281 agaatcggtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5341 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5401 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5461 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5521 tccatccagt ctatcaactg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5581 ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5641 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5701 aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca gaagtaagtt ggccgcagtg
     5761 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5821 tgcttttctg tgactggtga atactcaacc aagtcattct gagaatagtg tatgcggcga
     5881 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5941 aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6001 ttgagatcca gttcgatgta acccactcgc gcacccaact gatcttcagc atcttttact
     6061 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6121 agggcgacac ggaaatgttg aatactcatg cgaaagcctc ctgctgatcc tgtctcttga
     6181 tagatgtttg caaaagctag gactcgatta tacgttgacc gatcagacta tcaatatcta
     6241 ggtttaaaca gatcggtacg ctgccgtgac caggcttgag tacaatatct gacgattgga
     6301 cttcctgggt cccaagctgg attccgactg gctgactgcc a
//
