LOCUS       sequence-295772-        8077 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 8077)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8077)
  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-m4134475
FEATURES             Location/Qualifiers
     source          1..8077
                     /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             1952..1969
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2046..2167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2057..2076)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2111..2130
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2282..2328
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2358..2494
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2539..2610
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2836..3424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2916..2935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3924..5133
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4970..4989
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5137..5736
                     /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(5137..5156)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5633..5653
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5763..5811
                     /note="synthetic polyadenylation signal"
     CDS             join(6077..6145,6146..6937)
                     /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(6295..6314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6950..7086
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7231..7302
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7307..7353)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7371..7419
                     /note="synthetic polyadenylation signal"
     enhancer        7500..7803
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7805..8007
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7958..7978
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8076..8077,1..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 tcgatgagtg acaaaatcat
     1621 tcatctaacc gatgactcct ttgatacaga cgtgttgaag gccgatggtg cgatactggt
     1681 agatttctgg gctgaatggt gtggcccgtg caagatgatc gcacccattc tggacgagat
     1741 cgccgacgag tatcaaggaa aattaactgt cgctaagctt aatatagatc agaacccagg
     1801 gacggcacct aaatacggca tcaggggaat acctaccctc ctgctcttta agaatgggga
     1861 agttgccgcg acaaaagtgg gagccctgtc taagggccag ctaaaagagt tcttggacgc
     1921 taacctggca ggttcaggct cggggcacat gcaccatcac catcaccata gcagcgggct
     1981 cgtgccacgc ggctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     2041 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2101 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2161 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2221 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     2281 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     2341 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     2401 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     2461 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     2521 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     2581 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     2641 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     2701 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     2761 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2821 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2881 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2941 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3001 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3061 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3121 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3181 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3241 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3301 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3361 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3421 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3481 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3541 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3601 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3661 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     3721 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     3781 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     3841 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     3901 aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc
     3961 ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct
     4021 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     4081 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     4141 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     4201 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     4261 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     4321 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga
     4381 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct
     4441 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag
     4501 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa
     4561 gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc
     4621 tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     4681 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     4741 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     4801 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     4861 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     4921 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     4981 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     5041 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct
     5101 aaattctggc cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg
     5161 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     5221 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     5281 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5341 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5401 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5461 atggagggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5521 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5581 gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5641 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5701 cgcacctggt gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag
     5761 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct
     5821 aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     5881 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     5941 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     6001 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     6061 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6121 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6181 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6241 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     6301 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6361 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     6421 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6481 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6541 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6601 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     6661 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     6721 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6781 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6841 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6901 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     6961 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     7021 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     7081 ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
     7141 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
     7201 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
     7261 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
     7321 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
     7381 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
     7441 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7501 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7561 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7621 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7681 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7741 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7801 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     7861 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     7921 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     7981 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
     8041 atcctaccat ccactcgaca cacccgccag cggccgc
//
