LOCUS       sequence-318501-        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"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2167..2187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2188..2214
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2267..2388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2278..2297)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2332..2351
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2503..2549
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2579..2715
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2760..2831
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3057..3645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3137..3156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4391..4459,4460..5251)
                     /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(4609..4628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5264..5400
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5552..5623
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5628..5674)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5692..5740
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggccga aaatgtcact
      601 ggcttattta aggattgcag taaagtgatc acagggttgc acccgaccca agcgcctacg
      661 catctttcag tagacaccaa gttcaaaact gagggactct gtgtggacat accaggtatt
      721 cccaaggata tgacatacag acgcctgatc tcgatgatgg gcttcaagat gaactatcag
      781 gttaacggtt accccaatat gtttattacc cgagaggaag ctatccggca cgtgcgtgcc
      841 tggatagggt ttgacgtcga gggctgtcat gcaactaggg aagctgtagg aacaaatctg
      901 ccactgcaac tcggcttctc taccggagtc aacctagttg ccgtgcctac cgggtatgtg
      961 gatacaccaa acaatactga ttttagcaga gtctccgcaa aaccccctcc cggggaccag
     1021 ttcaaacact tgatccccct gatgtacaag ggtttaccat ggaacgtggt gcggattaaa
     1081 atagttcaga tgctcagcga cacactgaag aatttaagtg atagagttgt gtttgtactt
     1141 tgggcccatg gcttcgagct gacctctatg aagtatttcg ttaaaattgg acccgaacga
     1201 acgtgctgcc tgtgtgaccg gagggcgacg tgtttctcaa ctgcttccga tacctatgcc
     1261 tgctggcacc atagcatcgg ctttgattac gtgtacaacc cttttatgat tgacgtgcag
     1321 cagtggggat tcactgggaa cctacagtcg aatcacgatc tttactgtca ggtccatggg
     1381 aacgcacacg ttgctagctg cgacgcaatc atgacacgtt gccttgctgt ccacgagtgt
     1441 tttgtgaaac gagtggactg gactattgag tatcctatca ttggcgacga actgaagata
     1501 aatgcggctt gtaggaaggt gcagcatatg gtcgtcaaag ccgccctctt agctgataag
     1561 ttcccggtcc tgcacgatat tggcaatcca aaggccatca aatgcgtacc ccaggcagat
     1621 gttgagtgga agttttacga cgcgcaacca tgctccgata aggcctataa aatcgaagag
     1681 ctcttctaca gttatgccac ccattctgac aagtttaccg acggtgtgtg tttgttttgg
     1741 aattgcaacg tggatcgcta ccctgcaaac tctatcgttt gtcgcttcga cacacgcgta
     1801 ttgtccaatc tgaatctacc cggatgcgac ggaggttcac tgtatgtgaa caagcacgcc
     1861 ttccacaccc ccgcattcga taaatccgcc tttgtcaacc ttaaacaatt acctttcttt
     1921 tactacagcg actctccttg cgaatcccat ggaaagcaag tagtgagtga tatcgattac
     1981 gtcccactca aaagtgcaac atgtattacg agatgcaatc tcggtggtgc agtctgtaga
     2041 caccatgcca atgaatatcg gctgtatttg gacgcctata acatgatgat ttcagctggc
     2101 ttctctcttt gggtttacaa gcagtttgac acgtataatc tgtggaacac ctttacaagg
     2161 ctgcaagaaa acctttactt tcagtcgtac ccctatgacg taccagatta tgcgtaagct
     2221 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     2281 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2341 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2401 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2461 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     2521 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     2581 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     2641 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     2701 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     2761 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     2821 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     2881 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     2941 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     3001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3841 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     3901 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     3961 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     4021 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     4081 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     4141 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     4201 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     4261 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     4321 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     4381 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4501 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4561 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4621 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4681 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     4741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4801 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4921 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     4981 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     5041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5221 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     5281 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     5341 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     5401 gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     5461 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     5521 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     5581 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     5641 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     5701 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     5761 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
