LOCUS       sequence-295676-        6183 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 6183)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6183)
  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-m4134088
FEATURES             Location/Qualifiers
     source          1..6183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1589..1609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1958..1975
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2052..2173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2063..2082)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2117..2136
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2288..2334
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2364..2500
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2545..2616
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4176..4244,4245..5036)
                     /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(4394..4413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5049..5185
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5337..5408
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5413..5459)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5477..5525
                     /note="synthetic polyadenylation signal"
     enhancer        5606..5909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5911..6113
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6064..6084
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6182..6183,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcc gaaaatgtca ctggcttatt taaggattgc agtaaagtga tcacagggtt
       61 gcacccgacc caagcgccta cgcatctttc agtagacacc aagttcaaaa ctgagggact
      121 ctgtgtggac ataccaggta ttcccaagga tatgacatac agacgcctga tctcgatgat
      181 gggcttcaag atgaactatc aggttaacgg ttaccccaat atgtttatta cccgagagga
      241 agctatccgg cacgtgcgtg cctggatagg gtttgacgtc gagggctgtc atgcaactag
      301 ggaagctgta ggaacaaatc tgccactgca actcggcttc tctaccggag tcaacctagt
      361 tgccgtgcct accgggtatg tggatacacc aaacaatact gattttagca gagtctccgc
      421 aaaaccccct cccggggacc agttcaaaca cttgatcccc ctgatgtaca agggtttacc
      481 atggaacgtg gtgcggatta aaatagttca gatgctcagc gacacactga agaatttaag
      541 tgatagagtt gtgtttgtac tttgggccca tggcttcgag ctgacctcta tgaagtattt
      601 cgttaaaatt ggacccgaac gaacgtgctg cctgtgtgac cggagggcga cgtgtttctc
      661 aactgcttcc gatacctatg cctgctggca ccatagcatc ggctttgatt acgtgtacaa
      721 cccttttatg attgacgtgc agcagtgggg attcactggg aacctacagt cgaatcacga
      781 tctttactgt caggtccatg ggaacgcaca cgttgctagc tgcgacgcaa tcatgacacg
      841 ttgccttgct gtccacgagt gttttgtgaa acgagtggac tggactattg agtatcctat
      901 cattggcgac gaactgaaga taaatgcggc ttgtaggaag gtgcagcata tggtcgtcaa
      961 agccgccctc ttagctgata agttcccggt cctgcacgat attggcaatc caaaggccat
     1021 caaatgcgta ccccaggcag atgttgagtg gaagttttac gacgcgcaac catgctccga
     1081 taaggcctat aaaatcgaag agctcttcta cagttatgcc acccattctg acaagtttac
     1141 cgacggtgtg tgtttgtttt ggaattgcaa cgtggatcgc taccctgcaa actctatcgt
     1201 ttgtcgcttc gacacacgcg tattgtccaa tctgaatcta cccggatgcg acggaggttc
     1261 actgtatgtg aacaagcacg ccttccacac ccccgcattc gataaatccg cctttgtcaa
     1321 ccttaaacaa ttacctttct tttactacag cgactctcct tgcgaatccc atggaaagca
     1381 agtagtgagt gatatcgatt acgtcccact caaaagtgca acatgtatta cgagatgcaa
     1441 tctcggtggt gcagtctgta gacaccatgc caatgaatat cggctgtatt tggacgccta
     1501 taacatgatg atttcagctg gcttctctct ttgggtttac aagcagtttg acacgtataa
     1561 tctgtggaac acctttacaa ggctgcaaga aaacctttac tttcagtcga tgagtgacaa
     1621 aatcattcat ctaaccgatg actcctttga tacagacgtg ttgaaggccg atggtgcgat
     1681 actggtagat ttctgggctg aatggtgtgg cccgtgcaag atgatcgcac ccattctgga
     1741 cgagatcgcc gacgagtatc aaggaaaatt aactgtcgct aagcttaata tagatcagaa
     1801 cccagggacg gcacctaaat acggcatcag gggaatacct accctcctgc tctttaagaa
     1861 tggggaagtt gccgcgacaa aagtgggagc cctgtctaag ggccagctaa aagagttctt
     1921 ggacgctaac ctggcaggtt caggctcggg gcacatgcac catcaccatc accatagcag
     1981 cgggctcgtg ccacgcggct aagctagctt gactgactga gatacagcgt accttcagct
     2041 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2101 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2161 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2221 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     2281 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     2341 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     2401 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     2461 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     2521 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     2581 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     2641 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     2701 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     2761 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     3721 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     3781 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     3841 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     3901 ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     3961 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     4021 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     4081 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     4141 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     4201 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4261 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     4321 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     4381 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4441 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     4501 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4561 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     4621 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4681 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     4741 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     4801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4861 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     5041 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     5101 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     5161 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     5221 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     5281 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     5341 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     5401 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     5461 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     5521 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     5581 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5641 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     5701 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     5761 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     5821 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5881 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     5941 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     6001 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     6061 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     6121 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     6181 cgc
//
