LOCUS       sequence-300706-        6440 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 6440)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6440)
  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-m4252864
FEATURES             Location/Qualifiers
     source          1..6440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    26..74
                     /note="synthetic polyadenylation signal"
     terminator      92..138
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    143..214
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(366..502)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(515..1306,1307..1375))
                     /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     1138..1157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1641..1689
                     /note="synthetic polyadenylation signal"
     CDS             complement(1716..2315)
                     /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(1799..1819)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2296..2315
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2319..3528)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2463..2482)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4028..4616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4517..4536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4842..4913
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4958..5094)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5124..5170)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5285..5406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5322..5341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5376..5395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5459..5536)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             complement(5537..5557)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5878..6080)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5907..5927)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6082..6385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat ggatgtgtgt
       61 tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc ctctaaacgg
      121 gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac cactagaggg
      181 aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg ctgccaaaaa
      241 ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc gccacggctg
      301 ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt aaacgattcg
      361 ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca agttggggaa
      421 gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc gtcgcatcag
      481 gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc agtgaggcac
      541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc cgggaagcta
      781 gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct acaggcatcg
      841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccaatcg
      961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac tcaaccaagt
     1081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt tcttcggggc
     1201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgcgcac
     1261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatgcgaa
     1381 agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact cgattatacg
     1441 ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc cgtgaccagg
     1501 cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc cgactggctg
     1561 actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt ctcggtacac
     1621 caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     1681 gattttatta tctaagacta ggcttacata ctagcctagg caccgggctt gcgggtcatg
     1741 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     1801 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
     1861 tcggcggcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     1921 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     1981 aggccctcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     2041 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     2101 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     2161 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     2221 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     2281 gtggcgaggc gcaccgtggg cttgtactcg gtcatgttgt ctaacaaaaa agccaaaaac
     2341 ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat tgacccaaat
     2401 gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa ccccgagcgc
     2461 atagttcaaa accggagctt cagctactta agaagatagg tacataaaac cgaccaaaga
     2521 aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg attcaggaga
     2581 gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg cacacctacc
     2641 ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac gacacgacga
     2701 gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata actgccgccc
     2761 ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc agggtcccca
     2821 gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga agggctcaag
     2881 accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct cacagcccgc
     2941 ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc gctcccccca
     3001 acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg cggtctctcc
     3061 acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca gcccagcagc
     3121 ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga cccaaatccc
     3181 ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca tcggcgttca
     3241 ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc ctcgcctgtt
     3301 ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc taaaatgtcc
     3361 ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc tgtcctgagc
     3421 gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca gcgctcgccg
     3481 tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag gcgcgccccg
     3541 atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt gaacttctct
     3601 cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata ggcgctgcca
     3661 agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc gagaggtagc
     3721 ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt ggagagttcg
     3781 gaaatttaaa tgcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3841 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3901 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3961 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4141 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4201 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4261 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4321 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4381 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4441 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4501 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4561 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4621 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgattt
     4681 aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg accagatcga acttgactga
     4741 cgcaattctg agcgtagtcg aaacccagac aagtcgatct cgccaagtag ctgacttgac
     4801 tggttaatta agtttcgata gcccaaggta accaatgaga ttagggaggg aagaaagttt
     4861 gatgaactac ttctgacccc tagtggtgtc cagaaaagac cattaaagga atgtcttgat
     4921 cgaaatcgag agatcgcctg caggatagct gacgacctta aggataaatt tctggtaagg
     4981 aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa tctggcatat
     5041 gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca atgccggtag
     5101 atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga ggccccaagg
     5161 ggttatgcta cgataccgat agagatgggc caataccaca tttgtagagg ttttacttgc
     5221 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     5281 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5401 atcttatcat gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag ctagcttaga
     5461 gggcaccaga cgaggatatc tttttgaatc tgttggctgc actcaccgcg ataaaattct
     5521 ttttccatcg cctcttcgac tgaaagtaaa ggttttctaa ggttgccctg ttgtccagca
     5581 tttcttcaca aagcttgttt atgtctacag caccctgcat ggaaagcaaa acagaaagta
     5641 gtgaaaccat tttttcaaag gcttcagtag tatctttagc taagagaatg tcattgtgta
     5701 actggacaca ttgagcccac aatttagatg atgattctac tctgagttgt tgcaaaactg
     5761 agagtaagac tactgatgtg cactttacat ctgacatttt catggtggcg gccgctggcg
     5821 ggtgtgtcga gtggatggta ggatcgacaa agatctgatc tgacggttca ctaaaccagc
     5881 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     5941 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     6001 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     6061 gtactgccaa aaccgcatca gcatggtaat agcgatgact aatacgtaga tgtactgcca
     6121 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     6181 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     6241 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     6301 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     6361 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     6421 cgtaattgat tactagatct
//
