LOCUS       sequence-295104-        4500 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 4500)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4500)
  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-m4133989
FEATURES             Location/Qualifiers
     source          1..4500
                     /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"
     CDS             join(2493..2561,2562..3353)
                     /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(2711..2730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3366..3502
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3654..3725
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3730..3776)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3794..3842
                     /note="synthetic polyadenylation signal"
     enhancer        3923..4226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4228..4430
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4381..4401
                     /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 gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     2281 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     2341 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     2401 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     2461 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     2521 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2581 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     2641 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     2701 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2761 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     2821 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2881 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     2941 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3001 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     3061 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     3121 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3181 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3241 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3301 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     3361 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     3421 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     3481 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     3541 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     3601 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     3661 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     3721 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     3781 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     3841 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     3901 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3961 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4021 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4081 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4141 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4201 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     4261 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4321 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4381 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4441 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
