LOCUS       sequence-306087-        7184 bp    DNA     circular SYN 15-APR-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 7184)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7184)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134017
FEATURES             Location/Qualifiers
     source          1..7184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    65..136
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(141..187)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    205..253
                     /note="synthetic polyadenylation signal"
     enhancer        334..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     792..812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1957..1977
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1978..2004
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2057..2178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2068..2087)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2122..2141
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2293..2339
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2369..2505
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2550..2621
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2847..3435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2927..2946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3935..5144
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4981..5000
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5148..5747
                     /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(5148..5167)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5644..5664
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5774..5822
                     /note="synthetic polyadenylation signal"
     CDS             join(6088..6156,6157..6948)
                     /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(6306..6325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6961..7097
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
       61 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
      121 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
      181 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
      241 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
      661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      841 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
      901 ccagcggccg ccaccatgtc actggagaac gttgctttca atgtagtgaa caaggggcac
      961 tttgacggac agcaaggcga agtgccggtc agtatcataa ataatacggt gtacaccaaa
     1021 gtcgatggtg ttgacgtgga gttatttgaa aacaagacaa ctttgcctgt caatgtggcc
     1081 ttcgagctgt gggcaaaacg caacattaag ccagtacccg aagtgaaaat cctcaacaat
     1141 cttggcgttg atattgccgc aaacactgtt atctgggatt ataagcgaga tgcgcctgcc
     1201 catatatcca ccattggagt gtgttctatg acagacatcg ctaaaaagcc caccgagaca
     1261 atttgcgcgc cactcacggt attctttgac gggagggtcg acggtcaggt ggatctgttt
     1321 agaaatgccc gtaacggcgt acttattact gaaggaagcg tgaaaggcct acagcccagt
     1381 gttgggccga agcaagcaag cctaaatgga gtcacactga tcggagaggc tgtgaaaacc
     1441 cagttcaact actataagaa ggtcgacggc gtggttcagc agctgcctga gacatacttt
     1501 acccaatcgc ggaatctgca agaatttaag cccagatccc agatggagat tgacttcttg
     1561 gaactggcca tggatgagtt catcgaacgg tataagctcg agggctacgc cttcgagcac
     1621 atagtttatg gggatttttc tcattcacag ttaggcggtc tgcacttgct catcggatta
     1681 gcgaaacgct ttaaagagag cccattcgaa cttgaagact ttatcccaat ggactcaact
     1741 gtgaagaatt acttcataac tgacgctcag acagggtcca gcaagtgcgt ctgttctgtt
     1801 attgatttgc tacttgatga tttcgtggag attattaaat cacaggacct gtccgtggtt
     1861 tcgaaggtcg tgaaagtaac catcgactat acggaaatta gtttcatgct ttggtgtaag
     1921 gatggccatg tggagacatt ttatcctaag ctccaggaaa acctttactt tcagtcgtac
     1981 ccctatgacg taccagatta tgcgtaagct agcttgactg actgagatac agcgtacctt
     2041 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2101 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2161 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2221 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     2281 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     2341 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     2401 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     2461 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     2521 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     2581 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     2641 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     2701 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     2761 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     2821 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2881 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2941 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3001 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3061 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3121 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3181 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3241 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3301 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3361 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3421 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3481 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3541 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3601 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3661 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     3721 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     3781 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     3841 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     3901 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     3961 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     4021 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     4081 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     4141 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     4201 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     4261 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     4321 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     4381 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     4441 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     4501 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     4561 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     4621 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     4681 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     4741 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     4801 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     4861 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     4921 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     4981 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     5041 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta
     5101 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca
     5161 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     5221 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     5281 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5341 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     5401 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     5461 cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     5521 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     5581 tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     5641 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     5701 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag
     5761 cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
     5821 tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     5881 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     5941 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     6001 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     6061 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     6121 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6181 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     6241 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     6301 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6361 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     6421 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6481 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     6541 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6601 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     6661 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     6721 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6781 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6841 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6901 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     6961 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     7021 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     7081 cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc
     7141 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctc
//
