LOCUS       sequence-300708-        6386 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 6386)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6386)
  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-m4252806
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    74..145
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(150..196)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    214..262
                     /note="synthetic polyadenylation signal"
     enhancer        343..646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        648..850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     801..821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1177..1197
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1198..1215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1268..1389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1279..1298)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1333..1352
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1504..1550
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1580..1716
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1761..1832
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2058..2646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2138..2157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3146..4355
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4192..4211
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4359..4958
                     /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(4359..4378)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4855..4875
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4985..5033
                     /note="synthetic polyadenylation signal"
     CDS             join(5299..5367,5368..6159)
                     /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(5517..5536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6172..6308
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
       61 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
      121 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
      181 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
      241 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
      301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
      661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
      901 acacacccgc cagcggccgc caccatgagc aaaatgtctg acgtcaagtg tacatccgta
      961 gttctgttat cagtgcttca acagctgcgg gtggagagtt cgtcaaagct ctgggcccag
     1021 tgcgtgcagc tgcacaatga tatattgctc gcgaaagata ctacggaagc tttcgagaag
     1081 atggtcagct tgctatctgt tctactgagc atgcaaggag cagtagacat caacaaactt
     1141 tgcgaagaga tgttagacaa ccgagccacc ctgcaagaaa acctttactt tcagtcgcac
     1201 catcaccatc accattaagc tagcttgact gactgagata cagcgtacct tcagctcaca
     1261 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     1321 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1381 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1441 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     1501 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     1561 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     1621 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     1681 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga
     1741 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     1801 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa
     1861 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     1921 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     1981 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2221 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2461 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2881 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg
     2941 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc
     3001 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga
     3061 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga
     3121 ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc
     3181 gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc
     3241 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
     3301 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
     3361 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
     3421 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
     3481 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
     3541 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
     3601 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
     3661 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
     3721 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
     3781 aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
     3841 gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg
     3901 tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg
     3961 ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt
     4021 ggggccacca gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg
     4081 gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag
     4141 tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc
     4201 ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt
     4261 ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg
     4321 ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac aagcccacgg
     4381 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4441 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4501 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4561 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4621 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4681 agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4741 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4801 tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4861 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4921 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt
     4981 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     5041 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     5101 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     5161 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     5221 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     5281 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5341 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5401 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5461 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     5521 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5581 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     5641 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5701 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5761 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5821 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     5881 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     5941 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6001 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6061 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6121 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     6181 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     6241 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     6301 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     6361 gagcttacgc gaacagccgt ggcgct
//
