LOCUS       sequence-320324-        7769 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 7769)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7769)
  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-m4133659
FEATURES             Location/Qualifiers
     source          1..7769
                     /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             2191..2256
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2309..2430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2320..2339)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2374..2393
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2545..2591
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2621..2757
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2802..2873
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3099..3687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3179..3198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4187..5396
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5233..5252
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5400..5999
                     /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(5400..5419)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5896..5916
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6026..6074
                     /note="synthetic polyadenylation signal"
     CDS             join(6340..6408,6409..7200)
                     /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(6558..6577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7213..7349
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7501..7572
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7577..7623)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7641..7689
                     /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 tcagtcgatg gattataagg accacgatgg agactacaaa
     2221 gatcatgaca ttgattacaa ggacgacgat gacaaataag ctagcttgac tgactgagat
     2281 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     2341 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2401 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2461 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2521 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     2581 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     2641 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     2701 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     2761 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     2821 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     2881 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     2941 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     3001 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     3061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3901 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     3961 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     4021 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     4081 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     4141 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg
     4201 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa
     4261 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
     4321 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
     4381 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
     4441 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
     4501 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
     4561 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
     4621 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     4681 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     4741 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     4801 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa
     4861 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca
     4921 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg
     4981 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc
     5041 tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc
     5101 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc
     5161 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc
     5221 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt
     5281 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt
     5341 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca
     5401 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     5461 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     5521 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5581 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5641 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5701 cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg cccaaggagc
     5761 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5821 gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg
     5881 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5941 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg
     6001 ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt
     6061 tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg
     6121 cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac
     6181 ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg
     6241 tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca
     6301 aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt
     6361 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6421 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg
     6481 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg
     6541 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6601 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca
     6661 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6721 agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc
     6781 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6841 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc
     6901 ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac
     6961 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7021 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7081 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7141 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7201 ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     7261 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     7321 tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct
     7381 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     7441 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     7501 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     7561 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     7621 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     7681 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     7741 ttagttcata gcccatatat ggagttccg
//
