LOCUS       sequence-301667-        7203 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 7203)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7203)
  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-m4252952
FEATURES             Location/Qualifiers
     source          1..7203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      50..96
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    101..172
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(324..460)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(473..1264,1265..1333))
                     /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     1096..1115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1599..1647
                     /note="synthetic polyadenylation signal"
     CDS             complement(1674..2702)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2706..3915)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2850..2869)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4415..5003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4904..4923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5229..5300
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5345..5481)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5511..5557)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5672..5793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5709..5728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5763..5782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5886..5903)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(6252..6272)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6599..6801)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6628..6648)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6803..7106
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(7187..7203,1..32)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat agcataaccc
       61 cttggggcct ctaaacgggt cttgaggggt tttttgaatt gagaggtagc tgaagctgct
      121 gttatgacca ctagagggaa gaagatacct gtggagctaa tggtccaaga tgtcgggagc
      181 cgcgatcgct gccaaaaagg taagctgacg atagctgacg agattcgatg cccgacgagc
      241 aaatgagcgc cacggctgtt cgcgtaagct cgaaacgacg gagattcgat agagccagag
      301 tcgagtttaa acgattcggg atcttaagga taaatttctg gtaaggagga cacgtatgga
      361 agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg gagtataaga
      421 cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc acctagaatc ggttaccaat
      481 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      541 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      601 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
      661 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatca
      721 actgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttacgc aaggttgttg
      781 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      841 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      901 ccttcggtcc tccaatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
      961 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1021 gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1081 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1141 gaaagcgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1201 tgtaacccac tcgcgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1261 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1321 gttgaatact catgcgaaag cctcctgctg atcctgtctc ttgatagatg tttgcaaaag
     1381 ctaggactcg attatacgtt gaccgatcag actatcaata tctaggttta aacagatcgg
     1441 tacgctgccg tgaccaggct tgagtacaat atctgacgat tggacttcct gggtcccaag
     1501 ctggattccg actggctgac tgccaagcaa cgatatggat cgtcgagact cgcactgaga
     1561 ggatcgttct cggtacacca ggcgcgccta gcgttacaca cacaaaaaac caacacacat
     1621 ccatcttcga tggatagcga ttttattatc taagactagg cttacatact agcttattcc
     1681 tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag
     1741 ccatcggtcc agacggcggc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct
     1801 ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg
     1861 tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgg
     1921 ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa
     1981 gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc
     2041 gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag
     2101 ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc
     2161 tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga
     2221 tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt
     2281 ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggcggc agcgatggca
     2341 tccattgcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc
     2401 aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct gaactcccca
     2461 atgtcaagca cttccggaat cgggagcgcg gcggatgcaa agtgccgata aacataacga
     2521 tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca
     2581 tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg
     2641 tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttaccc
     2701 atgttgtcta acaaaaaagc caaaaacggc cagaatttag cggacaattt acaagtctaa
     2761 cactgaaaat tacatattga cccaaatgat tacatttcaa aaggtgccta aaaaacttca
     2821 caaaacacac tcgccaaccc cgagcgcata gttcaaaacc ggagcttcag ctacttaaga
     2881 agataggtac ataaaaccga ccaaagaaac tgacgcctca cttatccctc ccctcaccag
     2941 aggtccggcg cctgtcgatt caggagagcc taccctaggc ccgaaccctg cgtcctgcga
     3001 cggagaaaag cctaccgcac acctaccggc tggtggcccc accctgcatt ataagccaac
     3061 agaacgggtg acgtcacgac acgacgaggg cgcgcgctcc caaaggtacg ggtgcactgc
     3121 ccaacggcac cgccataact gccgcccccg caacagacga caaaccgagt tctccagtca
     3181 gtgacaaact tcacgtcagg gtccccagat ggtgcccagc ccatctcacc cgaataagag
     3241 ctttcccgca ttagcgaagg gctcaagacc ttgggttctt gccgcccacc atgcccccca
     3301 ccttgtttca acgacctcac agcccgcctc acaagcgtct tccattcaag actcgggaac
     3361 agccgccatt ttgctgcgct ccccccaacc cccagttcag ggcaaccttg ctcgcggacc
     3421 cagactacag cccttggcgg tctctccaca cgcttccgtc ccaccgagcg gcccggcggc
     3481 cacgaaagcc ccggccagcc cagcagcccg ctactcacca agtgacgatc acagcgatcc
     3541 acaaacaaga accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc cacacccggc
     3601 gcgtccttat ataatcatcg gcgttcaccg ccccacggag atccctccgc agaatcgccg
     3661 agaagggact acttttcctc gcctgttccg ctctctggaa agaaaaccag tgccctagag
     3721 tcacccaagt cccgtcctaa aatgtccttc tgctgatact ggggttctaa ggccgagtct
     3781 tatgagcagc gggccgctgt cctgagcgtc cgggcggaag gatcaggacg ctcctgcgcc
     3841 cttcgtctga cgtggcagcg ctcgccgtga cgaggggggc gcccgcggga ggcgccaaaa
     3901 cccggcgcgg aggccaggcg cgccccgatt tcgatattcc tcggatctcg gatagatcgg
     3961 gattgaggat gtaggttgaa cttctctcgg gtgatcccga tctcggatat tcgggattgg
     4021 agtaatacct ggcgataggc gctgccaagc aatagccaag gcgcaaggcc ggccaagaga
     4081 cttgcgatag ttcgttcgag aggtagcctg caagatggat cggacgaaag gtttgaagat
     4141 tttccgacga gggccttgga gagttcggaa atttaaatgc aactatggat gaacgaaata
     4201 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4261 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4321 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4381 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4441 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4501 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4561 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4621 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4681 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4741 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4801 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4861 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4921 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4981 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5041 tttgctggcc ttttgctcac atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat
     5101 agcaaggacc agatcgaact tgactgacgc aattctgagc gtagtcgaaa cccagacaag
     5161 tcgatctcgc caagtagctg acttgactgg ttaattaagt ttcgatagcc caaggtaacc
     5221 aatgagatta gggagggaag aaagtttgat gaactacttc tgacccctag tggtgtccag
     5281 aaaagaccat taaaggaatg tcttgatcga aatcgagaga tcgcctgcag gatagctgac
     5341 gaccttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag
     5401 ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg
     5461 catttttttc tgcgccaatg ccggtagata gcaatccggc ccgaggggca caaaaaaccc
     5521 ctcaagaccc gtttagaggc cccaaggggt tatgctacga taccgataga gatgggccaa
     5581 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     5641 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5761 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtgagctga aggtacgctg
     5821 tatctcagtc agtcaagcta gcagaggcct gcggatcctt agccgcgtgg cacgagcccg
     5881 ctgctatggt gatggtgatg gtgcatgtgc cccgagcctg aacctgccag gttagcgtcc
     5941 aagaactctt ttagctggcc cttagacagg gctcccactt ttgtcgcggc aacttcccca
     6001 ttcttaaaga gcaggagggt aggtattccc ctgatgccgt atttaggtgc cgtccctggg
     6061 ttctgatcta tattaagctt agcgacagtt aattttcctt gatactcgtc ggcgatctcg
     6121 tccagaatgg gtgcgatcat cttgcacggg ccacaccatt cagcccagaa atctaccagt
     6181 atcgcaccat cggccttcaa cacgtctgta tcaaaggagt catcggttag atgaatgatt
     6241 ttgtcactca tcgactgaaa gtaaaggttt tcttgcaggg tggctcggtt gtctaacatc
     6301 tcttcgcaaa gtttgttgat gtctactgct ccttgcatgc tcagtagaac agatagcaag
     6361 ctgaccatct tctcgaaagc ttccgtagta tctttcgcga gcaatatatc attgtgcagc
     6421 tgcacgcact gggcccagag ctttgacgaa ctctccaccc gcagctgttg aagcactgat
     6481 aacagaacta cggatgtaca cttgacgtca gacattttgc tcatggtggc ggccgctggc
     6541 gggtgtgtcg agtggatggt aggatcgaca aagatctgat ctgacggttc actaaaccag
     6601 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     6661 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     6721 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     6781 tgtactgcca aaaccgcatc agcatggtaa tagcgatgac taatacgtag atgtactgcc
     6841 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     6901 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     6961 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     7021 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     7081 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     7141 ccgtaattga ttactagatc tggctacact cgatcagcag ttagataata aaatcgctat
     7201 cca
//
