LOCUS       Exported                8207 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133511
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8207)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8207
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1589..1609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1613..1678
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1747..1868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1758..1777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1812..1831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1983..2029
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2059..2195
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2240..2311
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2617..2636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3625..4834
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4671..4690
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4838..5866
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5893..5941
                     /note="synthetic polyadenylation signal"
     CDS             6207..7067
                     /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(6425..6444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7080..7216
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7361..7432
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7437..7483
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7501..7549
                     /note="synthetic polyadenylation signal"
     enhancer        7630..7933
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7935..8137
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8088..8108
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      8206..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggcc gaaaatgtca ctggcttatt taaggattgc agtaaagtga tcacagggtt
       61 gcacccgacc caagcgccta cgcatctttc agtagacacc aagttcaaaa ctgagggact
      121 ctgtgtggac ataccaggta ttcccaagga tatgacatac agacgcctga tctcgatgat
      181 gggcttcaag atgaactatc aggttaacgg ttaccccaat atgtttatta cccgagagga
      241 agctatccgg cacgtgcgtg cctggatagg gtttgacgtc gagggctgtc atgcaactag
      301 ggaagctgta ggaacaaatc tgccactgca actcggcttc tctaccggag tcaacctagt
      361 tgccgtgcct accgggtatg tggatacacc aaacaatact gattttagca gagtctccgc
      421 aaaaccccct cccggggacc agttcaaaca cttgatcccc ctgatgtaca agggtttacc
      481 atggaacgtg gtgcggatta aaatagttca gatgctcagc gacacactga agaatttaag
      541 tgatagagtt gtgtttgtac tttgggccca tggcttcgag ctgacctcta tgaagtattt
      601 cgttaaaatt ggacccgaac gaacgtgctg cctgtgtgac cggagggcga cgtgtttctc
      661 aactgcttcc gatacctatg cctgctggca ccatagcatc ggctttgatt acgtgtacaa
      721 cccttttatg attgacgtgc agcagtgggg attcactggg aacctacagt cgaatcacga
      781 tctttactgt caggtccatg ggaacgcaca cgttgctagc tgcgacgcaa tcatgacacg
      841 ttgccttgct gtccacgagt gttttgtgaa acgagtggac tggactattg agtatcctat
      901 cattggcgac gaactgaaga taaatgcggc ttgtaggaag gtgcagcata tggtcgtcaa
      961 agccgccctc ttagctgata agttcccggt cctgcacgat attggcaatc caaaggccat
     1021 caaatgcgta ccccaggcag atgttgagtg gaagttttac gacgcgcaac catgctccga
     1081 taaggcctat aaaatcgaag agctcttcta cagttatgcc acccattctg acaagtttac
     1141 cgacggtgtg tgtttgtttt ggaattgcaa cgtggatcgc taccctgcaa actctatcgt
     1201 ttgtcgcttc gacacacgcg tattgtccaa tctgaatcta cccggatgcg acggaggttc
     1261 actgtatgtg aacaagcacg ccttccacac ccccgcattc gataaatccg cctttgtcaa
     1321 ccttaaacaa ttacctttct tttactacag cgactctcct tgcgaatccc atggaaagca
     1381 agtagtgagt gatatcgatt acgtcccact caaaagtgca acatgtatta cgagatgcaa
     1441 tctcggtggt gcagtctgta gacaccatgc caatgaatat cggctgtatt tggacgccta
     1501 taacatgatg atttcagctg gcttctctct ttgggtttac aagcagtttg acacgtataa
     1561 tctgtggaac acctttacaa ggctgcaaga aaacctttac tttcagtcga tggattataa
     1621 ggaccacgat ggagactaca aagatcatga cattgattac aaggacgacg atgacaaata
     1681 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
     1741 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1801 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1861 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1921 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     1981 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     2041 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     2101 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     2161 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     2221 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     2281 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     2341 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     2401 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     2461 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3361 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     3421 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     3481 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     3541 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     3601 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     3661 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     3721 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     3781 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     3841 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     3901 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     3961 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     4021 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     4081 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     4141 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     4201 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     4261 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     4321 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     4381 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     4441 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     4501 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     4561 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     4621 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     4681 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     4741 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     4801 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     4861 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     4921 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     4981 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     5041 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     5101 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     5161 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     5221 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     5281 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     5341 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     5401 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     5461 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     5521 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     5581 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     5641 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     5701 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     5761 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     5821 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     5881 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     5941 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     6001 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     6061 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     6121 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     6181 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     6241 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6301 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     6361 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     6421 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6481 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     6541 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6601 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     6661 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6721 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     6781 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     6841 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6901 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6961 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7021 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     7081 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     7141 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     7201 agaaatttat ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     7261 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     7321 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     7381 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     7441 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     7501 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     7561 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     7621 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     7681 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7741 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7801 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     7861 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     7921 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     7981 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     8041 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     8101 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     8161 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
