LOCUS       sequence-295854-        6400 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 6400)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6400)
  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-m4133428
FEATURES             Location/Qualifiers
     source          1..6400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             272..316
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    369..490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(380..399)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     434..453
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      605..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      681..817
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    862..933
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1159..1747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1239..1258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2247..3456
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3293..3312
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3460..4059
                     /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(3460..3479)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3956..3976
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4086..4134
                     /note="synthetic polyadenylation signal"
     CDS             join(4400..4468,4469..5260)
                     /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(4618..4637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5273..5409
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5554..5625
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5630..5676)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5694..5742
                     /note="synthetic polyadenylation signal"
     enhancer        5823..6126
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6128..6330
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6281..6301
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc gaaagaaacc gcggcagcta agttcgagag
      301 gcagcacatg gacagttaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      361 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      421 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      481 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      541 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
      601 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
      661 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
      721 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
      781 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
      841 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
      901 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
      961 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     1021 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1081 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1861 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1921 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1981 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     2041 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2101 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2161 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2221 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     2281 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     2341 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     2401 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     2461 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     2521 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     2581 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     2641 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     2701 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     2761 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     2821 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     2881 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     2941 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     3001 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     3061 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     3121 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     3181 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     3241 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     3301 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     3361 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     3421 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     3481 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3541 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3601 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3661 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3721 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3781 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3841 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3901 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3961 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4021 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     4081 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     4141 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4201 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4261 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4321 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     4381 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     4441 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4501 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4561 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     4621 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4681 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     4741 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4801 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4861 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4921 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     4981 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     5041 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5101 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5161 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5221 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5281 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5341 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5401 tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     5461 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     5521 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     5581 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     5641 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     5701 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     5761 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     5821 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     5881 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     5941 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6001 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6061 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6121 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6181 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     6241 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     6301 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     6361 tcgatcctac catccactcg acacacccgc cagcggccgc
//
