LOCUS       sequence-295988-        5820 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 5820)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5820)
  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-m4133446
FEATURES             Location/Qualifiers
     source          1..5820
                     /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             1610..1636
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1689..1810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1700..1719)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1754..1773
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1925..1971
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2001..2137
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2182..2253
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2479..3067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2559..2578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3813..3881,3882..4673)
                     /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(4031..4050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4686..4822
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4974..5045
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5050..5096)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5114..5162
                     /note="synthetic polyadenylation signal"
     enhancer        5243..5546
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5548..5750
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5701..5721
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5819..5820,1..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 tttcagtcgt acccctatga
     1621 cgtaccagat tatgcgtaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
     1681 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1741 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1801 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1861 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1921 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1981 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     2041 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     2101 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     2161 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     2221 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     2281 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     2341 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     2401 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2641 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3121 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3181 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3241 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3301 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     3361 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     3421 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     3481 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     3541 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     3601 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     3661 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     3721 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     3781 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     3841 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3901 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     3961 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     4021 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4081 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     4141 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4201 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     4261 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4321 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     4381 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     4441 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4501 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4561 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4621 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     4681 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     4741 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     4801 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     4861 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     4921 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     4981 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     5041 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     5101 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     5161 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     5221 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     5281 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5341 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     5401 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     5461 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     5521 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     5581 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     5641 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5701 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     5761 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
