LOCUS       sequence-297404-        6395 bp    DNA     circular SYN 15-APR-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 6395)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6395)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4252809
FEATURES             Location/Qualifiers
     source          1..6395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             278..304
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    357..478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(368..387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     422..441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      593..639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      669..805
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    850..921
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1147..1735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1227..1246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2235..3444
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3281..3300
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3448..4047
                     /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(3448..3467)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3944..3964
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4074..4122
                     /note="synthetic polyadenylation signal"
     CDS             join(4388..4456,4457..5248)
                     /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(4606..4625)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5261..5397
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5549..5620
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5625..5671)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5689..5737
                     /note="synthetic polyadenylation signal"
     enhancer        5818..6121
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6123..6325
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6276..6296
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaagaaa acctttactt tcagtcgtac ccctatgacg taccagatta
      301 tgcgtaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      361 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      421 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      481 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      541 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      601 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
      661 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
      721 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
      781 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
      841 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
      901 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
      961 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1021 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1081 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1561 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1681 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1741 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1801 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1861 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1921 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     1981 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2041 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2101 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2161 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2221 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     2281 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     2341 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     2401 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     2461 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     2521 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     2581 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     2641 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     2701 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     2761 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     2821 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     2881 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     2941 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     3001 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     3061 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccaccag
     3121 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     3181 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     3241 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     3301 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     3361 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     3421 ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc
     3481 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3541 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     3601 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     3661 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     3721 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc
     3781 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     3841 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc
     3901 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     3961 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4021 tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa
     4081 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     4141 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     4201 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     4261 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     4321 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     4381 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4441 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4501 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4561 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4621 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4681 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4741 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4801 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4861 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4921 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     4981 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     5041 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5101 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5161 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5221 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     5281 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     5341 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     5401 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     5461 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     5521 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     5581 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     5641 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     5701 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     5761 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5821 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5881 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     5941 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6001 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     6061 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6121 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     6181 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     6241 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     6301 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     6361 cctaccatcc actcgacaca cccgccagcg gccgc
//
