LOCUS       sequence-295423-        5832 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 5832)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5832)
  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-m4134349
FEATURES             Location/Qualifiers
     source          1..5832
                     /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"
     CDS             join(3825..3893,3894..4685)
                     /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(4043..4062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4698..4834
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4986..5057
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5062..5108)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5126..5174
                     /note="synthetic polyadenylation signal"
     enhancer        5255..5558
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5560..5762
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5713..5733
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5831..5832,1..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 tgtaccgaga acgatcctct
     3601 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     3661 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     3721 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     3781 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     3841 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3901 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     3961 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     4021 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4081 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     4141 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4201 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     4261 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4321 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4381 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     4441 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4501 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4561 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4621 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4681 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     4741 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     4801 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     4861 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     4921 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     4981 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     5041 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     5101 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     5161 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     5221 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     5281 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5341 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5401 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5461 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5521 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     5581 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     5641 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5701 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5761 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     5821 gccagcggcc gc
//
