LOCUS       sequence-295821-        7732 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 7732)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7732)
  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-m4134433
FEATURES             Location/Qualifiers
     source          1..7732
                     /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             1604..1648
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1701..1822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1712..1731)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1766..1785
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1937..1983
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2013..2149
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2194..2265
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2491..3079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2571..2590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3579..4788
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4625..4644
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4792..5391
                     /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(4792..4811)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5288..5308
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5418..5466
                     /note="synthetic polyadenylation signal"
     CDS             join(5732..5800,5801..6592)
                     /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(5950..5969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6605..6741
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6886..6957
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6962..7008)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7026..7074
                     /note="synthetic polyadenylation signal"
     enhancer        7155..7458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7460..7662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7613..7633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7731..7732,1..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 tcgaaagaaa ccgcggcagc
     1621 taagttcgag aggcagcaca tggacagtta agctagcttg actgactgag atacagcgta
     1681 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1741 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1801 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1861 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     1921 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1981 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     2041 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     2101 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     2161 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     2221 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     2281 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     2341 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     2401 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     2461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3001 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3061 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3301 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     3361 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     3421 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     3481 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     3541 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     3601 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     3661 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     3721 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     3781 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     3841 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     3901 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     3961 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     4021 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     4081 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     4141 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     4201 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     4261 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     4321 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     4381 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     4441 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     4501 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     4561 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     4621 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     4681 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     4741 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     4801 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4861 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4921 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4981 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5041 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5101 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5161 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5221 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5281 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5341 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     5401 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     5461 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     5521 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     5581 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     5641 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     5701 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     5761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5821 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     5881 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     5941 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6001 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     6061 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6121 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     6181 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6241 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     6301 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     6361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6421 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6541 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     6601 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6661 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     6721 ttaccagaaa tttatcctta agatcgttta aactcgactc tggctctatc gaatctccgt
     6781 cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt
     6841 cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta
     6901 gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat
     6961 tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg
     7021 ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa
     7081 ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca
     7141 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     7201 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     7261 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     7321 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     7381 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     7441 gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     7501 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     7561 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     7621 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     7681 atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
