LOCUS       sequence-297888-        5313 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 5313)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5313)
  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-m4133294
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      226..362
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    514..585
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(590..636)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    654..702
                     /note="synthetic polyadenylation signal"
     enhancer        783..1086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1088..1290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1241..1261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2400..2420
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2424..2489
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2542..2663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2553..2572)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2607..2626
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2778..2824
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2854..2990
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3035..3106
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3332..3920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3412..3431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4666..4734,4735..5313,1..213)
                     /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(4884..4903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
       61 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      121 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
      181 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
      241 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
      301 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
      361 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
      421 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
      481 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
      541 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
      601 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
      661 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
      721 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      961 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1021 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1081 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     1141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     1201 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     1261 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     1321 tcgatcctac catccactcg acacacccgc cagcggccgc caccatgtta gaaaatgtgg
     1381 cttttaatgt tgtaaataag ggacactttg atggacaaca gggtgaagta ccagtttcta
     1441 tcattaataa cactgtttac acaaaagttg atggtgttga tgtagaattg tttgaaaata
     1501 aaacaacatt acctgttaat gtagcatttg agctttgggc taagcgcaac attaaaccag
     1561 taccagaggt gaaaatactc aataatttgg gtgtggacat tgctgctaat actgtgatct
     1621 gggactacaa aagagatgct ccagcacata tatctactat tggtgtttgt tctatgactg
     1681 acatagccaa gaaaccaact gaaacgattt gtgcaccact cactgtcttt tttgatggta
     1741 gagttgatgg tcaagtagac ttatttagaa atgcccgtaa tggtgttctt attacagaag
     1801 gtagtgttaa aggtttacaa ccatctgtag gtcccaaaca agctagtctt aatggagtca
     1861 cattaattgg agaagccgta aaaacacagt tcaattatta taagaaagtt gatggtgttg
     1921 tccaacaatt acctgaaact tactttactc agagtagaaa tttacaagaa tttaaaccca
     1981 ggagtcaaat ggaaattgat ttcttagaat tagctatgga tgaattcatt gaacggtata
     2041 aattagaagg ctatgccttc gaacatatcg tttatggaga ttttagtcat agtcagttag
     2101 gtggtttaca tctactgatt ggactagcta aacgttttaa ggaatcacct tttgaattag
     2161 aagattttat tcctatggac agtacagtta aaaactattt cataacagat gcgcaaacag
     2221 gttcatctaa gtgtgtgtgt tctgttattg atttattact tgatgatttt gttgaaataa
     2281 taaaatccca agatttatct gtagtttcta aggttgtcaa agtgactatt gactatacag
     2341 aaatttcatt tatgctttgg tgtaaagatg gccatgtaga aacattttac ccaaaattag
     2401 aaaaccttta ctttcagtcg atggattata aggaccacga tggagactac aaagatcatg
     2461 acattgatta caaggacgac gatgacaaat aagctagctt gactgactga gatacagcgt
     2521 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2581 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2641 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2701 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
     2761 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
     2821 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
     2881 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     2941 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
     3001 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
     3061 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
     3121 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
     3181 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
     3241 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
     3301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4081 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4141 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     4201 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     4261 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     4321 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     4381 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc
     4441 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     4501 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     4561 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     4621 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     4681 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4741 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     4801 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     4861 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4921 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     4981 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5041 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     5101 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5161 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5221 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     5281 atagactgga tggaggcgga taaagttgca gga
//
