LOCUS       sequence-301660-        6840 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 6840)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6840)
  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-m4252895
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        551..1760
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1597..1616
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1764..2792
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    2819..2867
                     /note="synthetic polyadenylation signal"
     CDS             join(3133..3201,3202..3993)
                     /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(3351..3370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4006..4142
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4294..4365
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4370..4416)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4434..4482
                     /note="synthetic polyadenylation signal"
     enhancer        4563..4866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4868..5070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5021..5041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5397..5417
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5418..5444
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5513..5634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5524..5543)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5578..5597
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5749..5795
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5825..5961
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6006..6077
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(6303..6840,1..51))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6383..6402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
       61 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg
      301 aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc
      361 tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg
      421 gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag
      481 agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg
      541 gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg
      601 gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc
      661 tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg
      721 acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa
      781 caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg
      841 aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg
      901 gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc
      961 tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg
     1021 agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg
     1081 gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc
     1141 gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct
     1201 tgagcccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg
     1261 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg
     1321 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc
     1381 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc caccagccgg
     1441 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc
     1501 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc
     1561 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc
     1621 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat
     1681 ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt
     1741 ttttggcttt tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg
     1801 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     1861 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     1921 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc
     1981 tcccgattcc ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct
     2041 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     2101 tgcagccggt cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg
     2161 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     2221 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     2281 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     2341 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     2401 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     2461 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     2521 ggcatccgga gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg
     2581 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     2641 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     2701 gaagcgccgc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     2761 gccccagcac tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa
     2821 taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc
     2881 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     2941 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     3001 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     3061 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     3121 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3181 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3241 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3301 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
     3361 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3421 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     3481 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3541 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3601 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3661 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
     3721 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
     3781 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3841 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3901 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3961 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     4021 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     4081 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     4141 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     4201 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     4261 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     4321 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     4381 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     4441 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     4501 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4561 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     4621 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4681 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4741 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4801 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4861 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     4921 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4981 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     5041 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     5101 tcgatcctac catccactcg acacacccgc cagcggccgc caccatgagc aaaatgtctg
     5161 acgtcaagtg tacatccgta gttctgttat cagtgcttca acagctgcgg gtggagagtt
     5221 cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga tatattgctc gcgaaagata
     5281 ctacggaagc tttcgagaag atggtcagct tgctatctgt tctactgagc atgcaaggag
     5341 cagtagacat caacaaactt tgcgaagaga tgttagacaa ccgagccacc ctgcaagaaa
     5401 acctttactt tcagtcgtac ccctatgacg taccagatta tgcgtaagga tccgcaggcc
     5461 tctgctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     5521 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5581 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     5641 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     5701 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     5761 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     5821 accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     5881 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     5941 ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat
     6001 caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     6061 actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt
     6121 caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc
     6181 agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa
     6241 atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
//
