LOCUS       sequence-295255-        5814 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 5814)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5814)
  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-m4133933
FEATURES             Location/Qualifiers
     source          1..5814
                     /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..1630
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1683..1804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1694..1713)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1748..1767
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1919..1965
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1995..2131
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2176..2247
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2473..3061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2553..2572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3807..3875,3876..4667)
                     /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(4025..4044)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4680..4816
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4968..5039
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5044..5090)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5108..5156
                     /note="synthetic polyadenylation signal"
     enhancer        5237..5540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5542..5744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5695..5715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5813..5814,1..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 tcgtacccct atgacgtacc
     1621 agattatgcg taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
     1681 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1741 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1801 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1861 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     1921 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     1981 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     2041 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     2101 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     2161 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     2221 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     2281 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     2341 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     2401 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2641 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2881 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     3301 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     3361 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     3421 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     3481 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     3541 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     3601 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     3661 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     3721 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     3781 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     3841 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3901 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     3961 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     4021 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4081 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     4141 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4201 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     4261 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4321 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     4381 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     4441 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4501 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4561 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4621 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     4681 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     4741 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     4801 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     4861 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     4921 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     4981 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     5041 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     5101 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     5161 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     5221 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     5281 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     5341 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     5401 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     5461 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     5521 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     5581 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     5641 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     5701 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     5761 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
