LOCUS       sequence-295765-        6177 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 6177)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6177)
  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-m4134382
FEATURES             Location/Qualifiers
     source          1..6177
                     /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"
     CDS             join(4170..4238,4239..5030)
                     /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(4388..4407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5043..5179
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5331..5402
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5407..5453)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5471..5519
                     /note="synthetic polyadenylation signal"
     enhancer        5600..5903
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5905..6107
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6058..6078
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6176..6177,1..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 cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     3961 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     4021 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     4081 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     4141 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     4201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4261 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     4321 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     4381 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4441 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     4501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4561 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     4621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4681 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     4741 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     4801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4861 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     5041 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     5101 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     5161 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     5221 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     5281 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     5341 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     5401 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     5461 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     5521 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     5581 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5641 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5701 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5761 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5821 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5881 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     5941 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6001 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     6061 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     6121 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
