LOCUS       sequence-295247-        7222 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 7222)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7222)
  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-m4134125
FEATURES             Location/Qualifiers
     source          1..7222
                     /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..1138
                     /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    1191..1312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1202..1221)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1256..1275
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1427..1473
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1503..1639
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1684..1755
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1981..2569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2061..2080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3069..4278
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4115..4134
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4282..4881
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4282..4301)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4778..4798
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4908..4956
                     /note="synthetic polyadenylation signal"
     CDS             join(5222..5290,5291..6082)
                     /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(5440..5459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6095..6231
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6376..6447
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6452..6498)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6516..6564
                     /note="synthetic polyadenylation signal"
     enhancer        6645..6948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6950..7152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7103..7123
                     /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 aagaggcgat ggaaaaagaa
     1081 ttttatcgcg gtgagtgcag ccaacagatt caaaaagata tcctcgtctg gtgccctcta
     1141 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     1201 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1321 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1381 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     1441 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     1501 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     1561 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     1621 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     1681 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     1741 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     1801 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1861 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     2821 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2881 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2941 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     3001 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     3061 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     3121 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     3181 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     3241 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     3301 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     3361 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     3421 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     3481 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     3541 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     3601 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     3661 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     3721 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     3781 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     3841 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     3901 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     3961 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     4021 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     4081 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     4141 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     4201 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     4261 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     4321 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4381 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4441 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4501 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4561 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     4621 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4681 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     4741 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4801 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4861 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     4921 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     4981 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     5041 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     5101 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     5161 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     5221 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5281 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5341 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5401 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5461 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5521 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5581 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5641 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5701 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5761 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     5821 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5881 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5941 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6001 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6061 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     6121 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     6181 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcgttta
     6241 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     6301 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     6361 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     6421 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     6481 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     6541 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     6601 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6661 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6721 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6781 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6841 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6901 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     6961 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     7021 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     7081 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     7141 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     7201 cgacacaccc gccagcggcc gc
//
