LOCUS       sequence-295575-        7616 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 7616)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7616)
  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-m4133927
FEATURES             Location/Qualifiers
     source          1..7616
                     /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             1061..1087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1156..1277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1167..1186)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1221..1240
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1392..1438
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1468..1604
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1649..1720
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1946..2534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2026..2045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3034..4243
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4080..4099
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4247..5275
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5302..5350
                     /note="synthetic polyadenylation signal"
     CDS             join(5616..5684,5685..6476)
                     /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(5834..5853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6489..6625
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6770..6841
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6846..6892)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6910..6958
                     /note="synthetic polyadenylation signal"
     enhancer        7039..7342
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7344..7546
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7497..7517
                     /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 tacccctatg acgtaccaga
     1081 ttatgcgtaa ggatccgcag gcctctgcta gcttgactga ctgagataca gcgtaccttc
     1141 agctcacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1201 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1261 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1321 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc
     1381 tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
     1441 cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg
     1501 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     1561 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct
     1621 gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact
     1681 aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg
     1741 ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt
     1801 tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt
     1861 ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc
     1921 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2461 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2521 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2581 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2641 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2701 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2761 ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct
     2821 ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg
     2881 ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat
     2941 atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg
     3001 agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc
     3061 ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg
     3121 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     3181 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     3241 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     3301 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     3361 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     3421 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     3481 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     3541 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     3601 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
     3661 tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg
     3721 tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     3781 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     3841 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     3901 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
     3961 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     4021 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     4081 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     4141 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa
     4201 attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc
     4261 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     4321 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     4381 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     4441 tcggcacttt gcatccgccg cgctcccgat tccggaagtg cttgacattg gggagttcag
     4501 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     4561 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc
     4621 cgccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     4681 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     4741 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     4801 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     4861 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     4921 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     4981 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc
     5041 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     5101 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     5161 cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg accgatggct gtgtagaagt
     5221 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataagctag
     5281 tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt
     5341 tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     5401 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     5461 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     5521 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     5581 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     5641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5701 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     5761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     5821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5881 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     5941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6001 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     6061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6121 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     6181 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     6241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     6481 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     6541 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     6601 ctccttacca gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct
     6661 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
     6721 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
     6781 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
     6841 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
     6901 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
     6961 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
     7021 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7081 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7141 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7201 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     7261 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     7321 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
     7381 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     7441 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     7501 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     7561 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
