LOCUS       Exported                8436 bp ds-DNA     circular SYN 17-MAR-2022
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4134256
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8436)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8436)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 17, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8436
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1811..1831
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1835..1900
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1969..2090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1980..1999)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2034..2053
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2205..2251
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2281..2417
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2462..2533
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2759..3347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2839..2858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3847..5056
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4893..4912
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5060..6088
                     /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    6115..6163
                     /note="synthetic polyadenylation signal"
     CDS             6429..7289
                     /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(6647..6666)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7302..7438
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7590..7661
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7666..7712
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7730..7778
                     /note="synthetic polyadenylation signal"
     enhancer        7859..8162
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8164..8366
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8317..8337
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      8435..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggcg gtgggggctt gcgtactatg taacagccag acgtcattgc gctgcggtgc
       61 atgtataaga aggccgtttc tgtgttgcaa atgctgctac gatcatgtta tttcgacttc
      121 tcacaagctt gtgctcagtg tcaatcctta tgtgtgtaat gcccccggat gcgacgtcac
      181 agatgttacc caactgtacc tgggcggcat gtcctattat tgtaagagcc acaaaccacc
      241 aatctccttc cccctgtgcg ccaacggaca ggtgtttggg ctctacaaaa acacatgtgt
      301 gggatctgac aatgtcaccg acttcaacgc tatcgccaca tgcgattgga ctaatgcagg
      361 cgactacatt ttagccaaca cctgtaccga acggctgaag ttatttgcgg ctgagactct
      421 aaaggcaact gaggaaacct tcaaactttc atatggtatc gccacggtac gtgaggtgtt
      481 gtctgatcga gaactgcatc tcagttggga ggtcgggaag cctagacctc cccttaatcg
      541 aaattacgtg ttcacaggtt atagggttac caagaacagc aaagtccaga tcggcgaata
      601 tacttttgag aagggagatt acggtgacgc cgtagtgtat cgtggcacaa caacatacaa
      661 actgaatgtt ggggattact tcgtgctgac gtcccacacc gttatgccgc tctcagctcc
      721 aactttggtt ccccaggaac attacgtgcg gattactgga ctctatccaa cgctaaacat
      781 aagcgacgag ttttccagca acgtggctaa ttaccaaaag gtcggcatgc agaagtactc
      841 cacccttcag ggtcctcctg ggactggcaa aagtcacttc gccatcggac tggcacttta
      901 ctatcccagt gcgcgcattg tatataccgc ctgctcacat gcggcagtgg acgccctgtg
      961 tgagaaggct ttaaagtatc tgccaataga caaatgctct cgcattatcc ctgccagggc
     1021 cagagtggag tgctttgata agtttaaagt gaactcgacc ctcgaacagt atgtcttctg
     1081 cacagttaac gcattgcccg aaacgaccgc cgatatagtt gtgttcgacg aaatctcaat
     1141 ggctaccaat tatgatctct ccgtcgtcaa tgcacggctg cgggccaaac actacgtcta
     1201 catcggggac cccgcgcagt taccagcacc gcgaacccta ctgactaagg gcacacttga
     1261 gccagagtac ttcaacagcg tatgtagact gatgaaaacc attggccctg atatgttcct
     1321 gggaacatgt cggaggtgcc ccgctgaaat agtggacacc gtttcagcac ttgtgtacga
     1381 caataagctc aaagcacaca aggataaatc cgcacaatgt tttaagatgt tttacaaagg
     1441 ggttattacc catgatgtgt ctagcgctat caatcgccct cagattggcg tggtgagaga
     1501 gtttttgacg cgaaatcccg cctggcgaaa ggccgtattc atcagtcctt acaactcgca
     1561 aaatgccgtc gctagtaaga ttttaggact ccctactcag acagttgata gctctcaagg
     1621 ttctgaatac gattatgtga tttttactca gactaccgaa acggcacatt cttgtaacgt
     1681 caaccgtttt aacgtggcca ttacaagggc caaggttggc atcttgtgta tcatgtcaga
     1741 ccgggacctg tatgataaac tgcaattcac atctttggag ataccgcgcc gcaatgtcgc
     1801 tacccttcag gaaaaccttt actttcagtc gatggattat aaggaccacg atggagacta
     1861 caaagatcat gacattgatt acaaggacga cgatgacaaa taaggatccg caggcctctg
     1921 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1981 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2041 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2101 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2161 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     2221 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     2281 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     2341 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     2401 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     2461 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     2521 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     2581 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2641 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2941 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3181 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3601 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3661 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3721 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3781 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3841 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3901 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3961 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     4021 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     4081 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     4141 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     4201 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     4261 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     4321 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     4381 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     4441 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     4501 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     4561 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     4621 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     4681 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     4741 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4801 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4861 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4921 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4981 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     5041 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     5101 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     5161 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     5221 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     5281 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     5341 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     5401 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     5461 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     5521 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     5581 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     5641 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     5701 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     5761 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5821 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     5881 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     5941 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     6001 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     6061 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     6121 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     6181 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     6241 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     6301 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     6361 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     6421 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6481 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6541 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6601 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6661 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6721 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6781 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6841 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6901 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6961 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     7021 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     7081 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7141 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7201 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7261 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     7321 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     7381 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     7441 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     7501 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     7561 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     7621 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     7681 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     7741 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     7801 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7861 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7921 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7981 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     8041 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     8101 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     8161 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     8221 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     8281 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     8341 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     8401 tcctaccatc cactcgacac acccgccagc ggccgc
//
