LOCUS       sequence-295235-        4851 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 4851)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4851)
  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-m4133539
FEATURES             Location/Qualifiers
     source          1..4851
                     /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             626..643
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    720..841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(731..750)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     785..804
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      956..1002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1032..1168
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1213..1284
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1510..2098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1590..1609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2844..2912,2913..3704)
                     /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(3062..3081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3717..3853
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4005..4076
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4081..4127)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4145..4193
                     /note="synthetic polyadenylation signal"
     enhancer        4274..4577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4579..4781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4732..4752
                     /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 tcagtcgatg agtgacaaaa tcattcatct
      301 aaccgatgac tcctttgata cagacgtgtt gaaggccgat ggtgcgatac tggtagattt
      361 ctgggctgaa tggtgtggcc cgtgcaagat gatcgcaccc attctggacg agatcgccga
      421 cgagtatcaa ggaaaattaa ctgtcgctaa gcttaatata gatcagaacc cagggacggc
      481 acctaaatac ggcatcaggg gaatacctac cctcctgctc tttaagaatg gggaagttgc
      541 cgcgacaaaa gtgggagccc tgtctaaggg ccagctaaaa gagttcttgg acgctaacct
      601 ggcaggttca ggctcggggc acatgcacca tcaccatcac catagcagcg ggctcgtgcc
      661 acgcggctaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
      721 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      781 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      841 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
      901 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
      961 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1021 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1081 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1141 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1201 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1261 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1321 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1381 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1441 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1501 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1561 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1621 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1681 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1741 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1801 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1861 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1921 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1981 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2041 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2101 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2161 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2221 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2281 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2341 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2401 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2461 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2521 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2581 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     2641 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     2701 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     2761 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     2821 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     2881 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2941 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     3001 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     3061 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3121 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     3181 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3241 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     3301 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3361 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     3421 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     3481 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3541 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3601 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3661 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     3721 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     3781 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     3841 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     3901 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     3961 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     4021 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     4081 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     4141 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     4201 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     4261 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4321 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4381 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4441 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4501 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4561 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4621 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4681 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4741 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     4801 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
