LOCUS       sequence-295134-        7673 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 7673)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7673)
  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-m4134360
FEATURES             Location/Qualifiers
     source          1..7673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1046..1066
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1067..1144
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1213..1334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1224..1243)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1278..1297
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1449..1495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1525..1661
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1706..1777
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2003..2591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2083..2102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3091..4300
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4137..4156
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4304..5332
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5359..5407
                     /note="synthetic polyadenylation signal"
     CDS             join(5673..5741,5742..6533)
                     /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(5891..5910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6546..6682
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6827..6898
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6903..6949)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6967..7015
                     /note="synthetic polyadenylation signal"
     enhancer        7096..7399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7401..7603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7554..7574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtca ctggagaacg ttgctttcaa tgtagtgaac aaggggcact ttgacggaca
       61 gcaaggcgaa gtgccggtca gtatcataaa taatacggtg tacaccaaag tcgatggtgt
      121 tgacgtggag ttatttgaaa acaagacaac tttgcctgtc aatgtggcct tcgagctgtg
      181 ggcaaaacgc aacattaagc cagtacccga agtgaaaatc ctcaacaatc ttggcgttga
      241 tattgccgca aacactgtta tctgggatta taagcgagat gcgcctgccc atatatccac
      301 cattggagtg tgttctatga cagacatcgc taaaaagccc accgagacaa tttgcgcgcc
      361 actcacggta ttctttgacg ggagggtcga cggtcaggtg gatctgttta gaaatgcccg
      421 taacggcgta cttattactg aaggaagcgt gaaaggccta cagcccagtg ttgggccgaa
      481 gcaagcaagc ctaaatggag tcacactgat cggagaggct gtgaaaaccc agttcaacta
      541 ctataagaag gtcgacggcg tggttcagca gctgcctgag acatacttta cccaatcgcg
      601 gaatctgcaa gaatttaagc ccagatccca gatggagatt gacttcttgg aactggccat
      661 ggatgagttc atcgaacggt ataagctcga gggctacgcc ttcgagcaca tagtttatgg
      721 ggatttttct cattcacagt taggcggtct gcacttgctc atcggattag cgaaacgctt
      781 taaagagagc ccattcgaac ttgaagactt tatcccaatg gactcaactg tgaagaatta
      841 cttcataact gacgctcaga cagggtccag caagtgcgtc tgttctgtta ttgatttgct
      901 acttgatgat ttcgtggaga ttattaaatc acaggacctg tccgtggttt cgaaggtcgt
      961 gaaagtaacc atcgactata cggaaattag tttcatgctt tggtgtaagg atggccatgt
     1021 ggagacattt tatcctaagc tccaggaaaa cctttacttt cagtcgaaga ggcgatggaa
     1081 aaagaatttt atcgcggtga gtgcagccaa cagattcaaa aagatatcct cgtctggtgc
     1141 cctctaagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
     1201 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1261 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1321 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1381 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
     1441 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
     1501 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
     1561 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     1621 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
     1681 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
     1741 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
     1801 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     1861 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     1921 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2821 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     2881 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     2941 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     3001 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     3061 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     3121 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     3181 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     3241 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     3301 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     3361 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     3421 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     3481 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     3541 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     3601 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     3661 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     3721 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     3781 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     3841 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3901 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3961 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     4021 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     4081 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     4141 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     4201 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     4261 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     4321 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     4381 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     4441 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     4501 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     4561 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4621 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     4681 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4741 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4801 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4861 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4921 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4981 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     5041 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     5101 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     5161 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     5221 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     5281 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     5341 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     5401 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     5461 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     5521 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     5581 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     5641 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     5701 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5761 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     5821 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     5881 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5941 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     6001 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6061 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     6121 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6181 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     6241 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     6301 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6361 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6421 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6481 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     6541 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     6601 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     6661 cttaccagaa atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg
     6721 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     6781 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     6841 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     6901 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     6961 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     7021 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     7081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     7141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     7201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     7261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     7321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     7381 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     7441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     7501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     7561 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     7621 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
