LOCUS       sequence-295448-        8083 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 8083)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8083)
  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-m4133994
FEATURES             Location/Qualifiers
     source          1..8083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1589..1609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1958..1975
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2052..2173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2063..2082)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2117..2136
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2288..2334
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2364..2500
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2545..2616
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3930..5139
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4976..4995
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5143..5742
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5143..5162)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5639..5659
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5769..5817
                     /note="synthetic polyadenylation signal"
     CDS             join(6083..6151,6152..6943)
                     /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(6301..6320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6956..7092
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7237..7308
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7313..7359)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7377..7425
                     /note="synthetic polyadenylation signal"
     enhancer        7506..7809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7811..8013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7964..7984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8082..8083,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcc gaaaatgtca ctggcttatt taaggattgc agtaaagtga tcacagggtt
       61 gcacccgacc caagcgccta cgcatctttc agtagacacc aagttcaaaa ctgagggact
      121 ctgtgtggac ataccaggta ttcccaagga tatgacatac agacgcctga tctcgatgat
      181 gggcttcaag atgaactatc aggttaacgg ttaccccaat atgtttatta cccgagagga
      241 agctatccgg cacgtgcgtg cctggatagg gtttgacgtc gagggctgtc atgcaactag
      301 ggaagctgta ggaacaaatc tgccactgca actcggcttc tctaccggag tcaacctagt
      361 tgccgtgcct accgggtatg tggatacacc aaacaatact gattttagca gagtctccgc
      421 aaaaccccct cccggggacc agttcaaaca cttgatcccc ctgatgtaca agggtttacc
      481 atggaacgtg gtgcggatta aaatagttca gatgctcagc gacacactga agaatttaag
      541 tgatagagtt gtgtttgtac tttgggccca tggcttcgag ctgacctcta tgaagtattt
      601 cgttaaaatt ggacccgaac gaacgtgctg cctgtgtgac cggagggcga cgtgtttctc
      661 aactgcttcc gatacctatg cctgctggca ccatagcatc ggctttgatt acgtgtacaa
      721 cccttttatg attgacgtgc agcagtgggg attcactggg aacctacagt cgaatcacga
      781 tctttactgt caggtccatg ggaacgcaca cgttgctagc tgcgacgcaa tcatgacacg
      841 ttgccttgct gtccacgagt gttttgtgaa acgagtggac tggactattg agtatcctat
      901 cattggcgac gaactgaaga taaatgcggc ttgtaggaag gtgcagcata tggtcgtcaa
      961 agccgccctc ttagctgata agttcccggt cctgcacgat attggcaatc caaaggccat
     1021 caaatgcgta ccccaggcag atgttgagtg gaagttttac gacgcgcaac catgctccga
     1081 taaggcctat aaaatcgaag agctcttcta cagttatgcc acccattctg acaagtttac
     1141 cgacggtgtg tgtttgtttt ggaattgcaa cgtggatcgc taccctgcaa actctatcgt
     1201 ttgtcgcttc gacacacgcg tattgtccaa tctgaatcta cccggatgcg acggaggttc
     1261 actgtatgtg aacaagcacg ccttccacac ccccgcattc gataaatccg cctttgtcaa
     1321 ccttaaacaa ttacctttct tttactacag cgactctcct tgcgaatccc atggaaagca
     1381 agtagtgagt gatatcgatt acgtcccact caaaagtgca acatgtatta cgagatgcaa
     1441 tctcggtggt gcagtctgta gacaccatgc caatgaatat cggctgtatt tggacgccta
     1501 taacatgatg atttcagctg gcttctctct ttgggtttac aagcagtttg acacgtataa
     1561 tctgtggaac acctttacaa ggctgcaaga aaacctttac tttcagtcga tgagtgacaa
     1621 aatcattcat ctaaccgatg actcctttga tacagacgtg ttgaaggccg atggtgcgat
     1681 actggtagat ttctgggctg aatggtgtgg cccgtgcaag atgatcgcac ccattctgga
     1741 cgagatcgcc gacgagtatc aaggaaaatt aactgtcgct aagcttaata tagatcagaa
     1801 cccagggacg gcacctaaat acggcatcag gggaatacct accctcctgc tctttaagaa
     1861 tggggaagtt gccgcgacaa aagtgggagc cctgtctaag ggccagctaa aagagttctt
     1921 ggacgctaac ctggcaggtt caggctcggg gcacatgcac catcaccatc accatagcag
     1981 cgggctcgtg ccacgcggct aagctagctt gactgactga gatacagcgt accttcagct
     2041 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2101 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2161 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2221 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     2281 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     2341 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     2401 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     2461 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     2521 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     2581 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     2641 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     2701 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     2761 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     3721 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     3781 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     3841 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     3901 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     3961 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     4021 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     4081 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     4141 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     4201 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     4261 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     4321 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     4381 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     4441 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     4501 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     4561 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     4621 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     4681 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     4741 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     4801 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     4861 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     4921 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     4981 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     5041 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     5101 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     5161 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     5221 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     5281 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     5341 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     5401 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5461 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5521 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5581 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5641 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5701 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     5761 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     5821 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     5881 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     5941 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     6001 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     6061 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6121 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6181 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     6241 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     6301 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6361 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     6421 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6481 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     6541 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6601 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     6661 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     6721 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6781 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6841 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6901 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     6961 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     7021 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     7081 atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     7141 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     7201 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     7261 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     7321 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     7381 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     7441 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     7501 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     7561 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     7621 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     7681 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     7741 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     7801 attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     7861 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     7921 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     7981 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct
     8041 ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
