LOCUS       sequence-295724-        8177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8177)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8177)
  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-m4133144
FEATURES             Location/Qualifiers
     source          1..8177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1583..1603
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1604..1648
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1717..1838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1728..1747)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1782..1801
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1953..1999
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2029..2165
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2210..2281
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2507..3095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2587..2606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3595..4804
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4641..4660
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4808..5836
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5863..5911
                     /note="synthetic polyadenylation signal"
     CDS             join(6177..6245,6246..7037)
                     /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(6395..6414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7050..7186
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7331..7402
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7407..7453)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7471..7519
                     /note="synthetic polyadenylation signal"
     enhancer        7600..7903
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7905..8107
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8058..8078
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8176..8177,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggaa aatgtaacag gactctttaa agattgtagt aaggtaatca ctgggttaca
       61 tcctacacag gcacctacac acctcagtgt tgacactaaa ttcaaaactg aaggtttatg
      121 tgttgacata cctggcatac ctaaggacat gacctataga agactcatct ctatgatggg
      181 ttttaaaatg aattatcaag ttaatggtta ccctaacatg tttatcaccc gcgaagaagc
      241 tataagacat gtacgtgcat ggattggctt cgatgtcgag gggtgtcatg ctactagaga
      301 agctgttggt accaatttac ctttacagct aggtttttct acaggtgtta acctagttgc
      361 tgtacctaca ggttatgttg atacacctaa taatacagat ttttccagag ttagtgctaa
      421 accaccgcct ggagatcaat ttaaacacct cataccactt atgtacaaag gacttccttg
      481 gaatgtagtg cgtataaaga ttgtacaaat gttaagtgac acacttaaaa atctctctga
      541 cagagtcgta tttgtcttat gggcacatgg ctttgagttg acatctatga agtattttgt
      601 gaaaatagga cctgagcgca cctgttgtct atgtgataga cgtgccacat gcttttccac
      661 tgcttcagac acttatgcct gttggcatca ttctattgga tttgattacg tctataatcc
      721 gtttatgatt gatgttcaac aatggggttt tacaggtaac ctacaaagca accatgatct
      781 gtattgtcaa gtccatggta atgcacatgt agctagttgt gatgcaatca tgactaggtg
      841 tctagctgtc cacgagtgct ttgttaagcg tgttgactgg actattgaat atcctataat
      901 tggtgatgaa ctgaagatta atgcggcttg tagaaaggtt caacacatgg ttgttaaagc
      961 tgcattatta gcagacaaat tcccagttct tcacgacatt ggtaacccta aagctattaa
     1021 gtgtgtacct caagctgatg tagaatggaa gttctatgat gcacagcctt gtagtgacaa
     1081 agcttataaa atagaagaat tattctattc ttatgccaca cattctgaca aattcacaga
     1141 tggtgtatgc ctattttgga attgcaatgt cgatagatat cctgctaatt ccattgtttg
     1201 tagatttgac actagagtgc tatctaacct taacttgcct ggttgtgatg gtggcagttt
     1261 gtatgtaaat aaacatgcat tccacacacc agcttttgat aaaagtgctt ttgttaattt
     1321 aaaacaatta ccatttttct attactctga cagtccatgt gagtctcatg gaaaacaagt
     1381 agtgtcagat atagattatg taccactaaa gtctgctacg tgtataacac gttgcaattt
     1441 aggtggtgct gtctgtagac atcatgctaa tgagtacaga ttgtatctcg atgcttataa
     1501 catgatgatc tcagctggct ttagcttgtg ggtttacaaa caatttgata cttataacct
     1561 ctggaacact tttacaagac ttgaaaacct ttactttcag tcgaaagaaa ccgcggcagc
     1621 taagttcgag aggcagcaca tggacagtta aggatccgca ggcctctgct agcttgactg
     1681 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac
     1741 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1801 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1861 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1921 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg
     1981 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag
     2041 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     2101 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     2161 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa
     2221 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct
     2281 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg
     2341 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg
     2401 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag
     2461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2941 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3301 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct
     3361 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac
     3421 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca
     3481 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca
     3541 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc
     3601 cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt
     3661 cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
     3721 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
     3781 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
     3841 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
     3901 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
     3961 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
     4021 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
     4081 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
     4141 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
     4201 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat
     4261 gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa
     4321 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     4381 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     4441 acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc
     4501 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     4561 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     4621 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     4681 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     4741 ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     4801 agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     4861 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     4921 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     4981 ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt
     5041 gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     5101 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     5161 ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg
     5221 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     5281 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     5341 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     5401 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     5461 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     5521 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     5581 aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     5641 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     5701 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg
     5761 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     5821 gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat
     5881 cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga
     5941 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     6001 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     6061 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     6121 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     6181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     6301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6361 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6481 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     6601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6721 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     6781 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7021 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     7081 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     7141 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc gtttaaactc
     7201 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     7261 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     7321 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     7381 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     7441 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     7501 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     7561 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     7621 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     7681 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     7741 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     7801 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     7861 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     7921 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     7981 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     8041 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     8101 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     8161 cacccgccag cggccgc
//
