LOCUS       sequence-295101-        5634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5634)
  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-m4134396
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1040..1060
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1409..1426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1503..1624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1514..1533)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1568..1587
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1739..1785
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1815..1951
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1996..2067
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2293..2881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2373..2392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3627..3695,3696..4487)
                     /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(3845..3864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4500..4636
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4788..4859
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4864..4910)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4928..4976
                     /note="synthetic polyadenylation signal"
     enhancer        5057..5360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5362..5564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5515..5535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtta gaaaatgtgg cttttaatgt tgtaaataag ggacactttg atggacaaca
       61 gggtgaagta ccagtttcta tcattaataa cactgtttac acaaaagttg atggtgttga
      121 tgtagaattg tttgaaaata aaacaacatt acctgttaat gtagcatttg agctttgggc
      181 taagcgcaac attaaaccag taccagaggt gaaaatactc aataatttgg gtgtggacat
      241 tgctgctaat actgtgatct gggactacaa aagagatgct ccagcacata tatctactat
      301 tggtgtttgt tctatgactg acatagccaa gaaaccaact gaaacgattt gtgcaccact
      361 cactgtcttt tttgatggta gagttgatgg tcaagtagac ttatttagaa atgcccgtaa
      421 tggtgttctt attacagaag gtagtgttaa aggtttacaa ccatctgtag gtcccaaaca
      481 agctagtctt aatggagtca cattaattgg agaagccgta aaaacacagt tcaattatta
      541 taagaaagtt gatggtgttg tccaacaatt acctgaaact tactttactc agagtagaaa
      601 tttacaagaa tttaaaccca ggagtcaaat ggaaattgat ttcttagaat tagctatgga
      661 tgaattcatt gaacggtata aattagaagg ctatgccttc gaacatatcg tttatggaga
      721 ttttagtcat agtcagttag gtggtttaca tctactgatt ggactagcta aacgttttaa
      781 ggaatcacct tttgaattag aagattttat tcctatggac agtacagtta aaaactattt
      841 cataacagat gcgcaaacag gttcatctaa gtgtgtgtgt tctgttattg atttattact
      901 tgatgatttt gttgaaataa taaaatccca agatttatct gtagtttcta aggttgtcaa
      961 agtgactatt gactatacag aaatttcatt tatgctttgg tgtaaagatg gccatgtaga
     1021 aacattttac ccaaaattag aaaaccttta ctttcagtcg atgagtgaca aaatcattca
     1081 tctaaccgat gactcctttg atacagacgt gttgaaggcc gatggtgcga tactggtaga
     1141 tttctgggct gaatggtgtg gcccgtgcaa gatgatcgca cccattctgg acgagatcgc
     1201 cgacgagtat caaggaaaat taactgtcgc taagcttaat atagatcaga acccagggac
     1261 ggcacctaaa tacggcatca ggggaatacc taccctcctg ctctttaaga atggggaagt
     1321 tgccgcgaca aaagtgggag ccctgtctaa gggccagcta aaagagttct tggacgctaa
     1381 cctggcaggt tcaggctcgg ggcacatgca ccatcaccat caccatagca gcgggctcgt
     1441 gccacgcggc taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
     1501 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1561 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1621 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1681 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     1741 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     1801 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1861 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     1921 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     1981 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     2041 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     2101 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     2161 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     2221 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3001 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3061 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     3121 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     3181 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     3241 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     3301 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     3361 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     3421 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     3481 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     3541 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     3601 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     3661 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3721 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     3781 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     3841 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3901 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     3961 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4021 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     4081 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4141 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     4201 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     4261 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4321 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4381 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4441 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     4501 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     4561 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     4621 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     4681 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     4741 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     4801 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     4861 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     4921 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     4981 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     5041 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     5101 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     5161 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     5221 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     5281 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     5341 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     5401 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     5461 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     5521 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     5581 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
