LOCUS       sequence-300724-        4479 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 4479)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4479)
  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-m4133277
FEATURES             Location/Qualifiers
     source          1..4479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      46..92
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    97..168
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(320..456)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(469..1260,1261..1329))
                     /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     1092..1111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2075..2663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2564..2583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2889..2960
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3005..3141)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3171..3217)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3332..3453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3369..3388)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3423..3442
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3506..3523)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3524..3544)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3871..4073)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3900..3920)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4075..4378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(4459..4479,1..28)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 agatggatgt gtgttggttt tttgtgtgtg taacgcaacg attgatagca taaccccttg
       61 gggcctctaa acgggtcttg aggggttttt tgaattgaga ggtagctgaa gctgctgtta
      121 tgaccactag agggaagaag atacctgtgg agctaatggt ccaagatgtc gggagccgcg
      181 atcgctgcca aaaaggtaag ctgacgatag ctgacgagat tcgatgcccg acgagcaaat
      241 gagcgccacg gctgttcgcg taagctcgaa acgacggaga ttcgatagag ccagagtcga
      301 gtttaaacga ttcgggatct taaggataaa tttctggtaa ggaggacacg tatggaagtg
      361 ggcaagttgg ggaagccgta tccgttgctg aatctggcat atgtgggagt ataagacgcg
      421 cagcgtcgca tcaggcattt ttttctgcgc caatgcacct agaatcggtt accaatgctt
      481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctatcaactg
      721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttacgcaagg ttgttgccat
      781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      901 cggtcctcca atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1021 atactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1141 gcgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1201 acccactcgc gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1321 aatactcatg cgaaagcctc ctgctgatcc tgtctcttga tagatgtttg caaaagctag
     1381 gactcgatta tacgttgacc gatcagacta tcaatatcta ggtttaaaca gatcggtacg
     1441 ctgccgtgac caggcttgag tacaatatct gacgattgga cttcctgggt cccaagctgg
     1501 attccgactg gctgactgcc aagcaacgat atggatcgtc gagactcgca ctgagaggat
     1561 cgttctcggt acaccaggcg cgccccgatt tcgatattcc tcggatctcg gatagatcgg
     1621 gattgaggat gtaggttgaa cttctctcgg gtgatcccga tctcggatat tcgggattgg
     1681 agtaatacct ggcgataggc gctgccaagc aatagccaag gcgcaaggcc ggccaagaga
     1741 cttgcgatag ttcgttcgag aggtagcctg caagatggat cggacgaaag gtttgaagat
     1801 tttccgacga gggccttgga gagttcggaa atttaaatgc aactatggat gaacgaaata
     1861 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     1921 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     1981 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2041 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2101 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2161 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2221 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2281 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2341 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2401 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2461 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2521 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2581 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2641 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2701 tttgctggcc ttttgctcac atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat
     2761 agcaaggacc agatcgaact tgactgacgc aattctgagc gtagtcgaaa cccagacaag
     2821 tcgatctcgc caagtagctg acttgactgg ttaattaagt ttcgatagcc caaggtaacc
     2881 aatgagatta gggagggaag aaagtttgat gaactacttc tgacccctag tggtgtccag
     2941 aaaagaccat taaaggaatg tcttgatcga aatcgagaga tcgcctgcag gatagctgac
     3001 gaccttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag
     3061 ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg
     3121 catttttttc tgcgccaatg ccggtagata gcaatccggc ccgaggggca caaaaaaccc
     3181 ctcaagaccc gtttagaggc cccaaggggt tatgctacga taccgataga gatgggccaa
     3241 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3301 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3361 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3421 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtgagctga aggtacgctg
     3481 tatctcagtc agtcaagcta gcttaatggt gatggtgatg gtgcgactga aagtaaaggt
     3541 tttcttgcag ggtggctcgg ttgtctaaca tctcttcgca aagtttgttg atgtctactg
     3601 ctccttgcat gctcagtaga acagatagca agctgaccat cttctcgaaa gcttccgtag
     3661 tatctttcgc gagcaatata tcattgtgca gctgcacgca ctgggcccag agctttgacg
     3721 aactctccac ccgcagctgt tgaagcactg ataacagaac tacggatgta cacttgacgt
     3781 cagacatttt gctcatggtg gcggccgctg gcgggtgtgt cgagtggatg gtaggatcga
     3841 caaagatctg atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac
     3901 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     3961 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     4021 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcagcatggt
     4081 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     4141 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     4201 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     4261 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     4321 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     4381 gaactccata tatgggctat gaactaatga ccccgtaatt gattactaga tctggctaca
     4441 ctcgatcagc agttagataa taaaatcgct atccatcga
//
