LOCUS       Exported                8432 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133152
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8432)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8432)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8432
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1805..1825
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1826..1903
                     /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    1972..2093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1983..2002)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2037..2056
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2208..2254
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2284..2420
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2465..2536
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2762..3350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2842..2861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3850..5059
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4896..4915
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5063..6091
                     /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    6118..6166
                     /note="synthetic polyadenylation signal"
     CDS             6432..7292
                     /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(6650..6669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7305..7441
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7586..7657
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7662..7708
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7726..7774
                     /note="synthetic polyadenylation signal"
     enhancer        7855..8158
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8160..8362
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8313..8333
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      8431..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggtt ggggcttgtg ttctttgcaa ttcacagact tcattaagat gtggtgcttg
       61 catacgtaga ccattcttat gttgtaaatg ctgttacgac catgtcatat caacatcaca
      121 taaattagtc ttgtctgtta atccgtatgt ttgcaatgct ccaggttgtg atgtcacaga
      181 tgtgactcaa ctttacttag gaggtatgag ctattattgt aaatcacata aaccacccat
      241 tagttttcca ttgtgtgcta atggacaagt ttttggttta tataaaaata catgtgttgg
      301 tagcgataat gttactgact ttaatgcaat tgcaacatgt gactggacaa atgctggtga
      361 ttacatttta gctaacacct gtactgaaag actcaagctt tttgcagcag aaacgctcaa
      421 agctactgag gagacattta aactgtctta tggtattgct actgtacgtg aagtgctgtc
      481 tgacagagaa ttacatcttt catgggaagt tggtaaacct agaccaccac ttaaccgaaa
      541 ttatgtcttt actggttatc gtgtaactaa aaacagtaaa gtacaaatag gagagtacac
      601 ctttgaaaaa ggtgactatg gtgatgctgt tgtttaccga ggtacaacaa cttacaaatt
      661 aaatgttggt gattattttg tgctgacatc acatacagta atgccattaa gtgcacctac
      721 actagtgcca caagagcact atgttagaat tactggctta tacccaacac tcaatatctc
      781 agatgagttt tctagcaatg ttgcaaatta tcaaaaggtt ggtatgcaaa agtattctac
      841 actccaggga ccacctggta ctggtaagag tcattttgct attggcctag ctctctacta
      901 cccttctgct cgcatagtgt atacagcttg ctctcatgcc gctgttgatg cactatgtga
      961 gaaggcatta aaatatttgc ctatagataa atgtagtaga attatacctg cacgtgctcg
     1021 tgtagagtgt tttgataaat tcaaagtgaa ttcaacatta gaacagtatg tcttttgtac
     1081 tgtaaatgca ttgcctgaga cgacagcaga tatagttgtc tttgatgaaa tttcaatggc
     1141 cacaaattat gatttgagtg ttgtcaatgc cagattacgt gctaagcact atgtgtacat
     1201 tggcgaccct gctcaattac ctgcaccacg cacattgcta actaagggca cactagaacc
     1261 agaatatttc aattcagtgt gtagacttat gaaaactata ggtccagaca tgttcctcgg
     1321 aacttgtcgg cgttgtcctg ctgaaattgt tgacactgtg agtgctttgg tttatgataa
     1381 taagcttaaa gcacataaag acaaatcagc tcaatgcttt aaaatgtttt ataagggtgt
     1441 tatcacgcat gatgtttcat ctgcaattaa caggccacaa ataggcgtgg taagagaatt
     1501 ccttacacgt aaccctgctt ggagaaaagc tgtctttatt tcaccttata attcacagaa
     1561 tgctgtagcc tcaaagattt tgggactacc aactcaaact gttgattcat cacagggctc
     1621 agaatatgac tatgtcatat tcactcaaac cactgaaaca gctcactctt gtaatgtaaa
     1681 cagatttaat gttgctatta ccagagcaaa agtaggcata ctttgcataa tgtctgatag
     1741 agacctttat gacaagttgc aatttacaag tcttgaaatt ccacgtagga atgtggcaac
     1801 tttagaaaac ctttactttc agtcgaagag gcgatggaaa aagaatttta tcgcggtgag
     1861 tgcagccaac agattcaaaa agatatcctc gtctggtgcc ctctaaggat ccgcaggcct
     1921 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     1981 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2041 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2101 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2161 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     2221 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     2281 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     2341 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     2401 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     2461 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     2521 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     2581 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     2641 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     2701 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2761 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2821 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2881 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2941 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3001 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3061 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3121 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3181 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3241 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3301 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3361 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3421 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3481 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     3601 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     3661 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     3721 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     3781 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     3841 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     3901 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     3961 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     4021 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     4081 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     4141 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     4201 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     4261 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     4321 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     4381 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     4441 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     4501 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     4561 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     4621 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     4681 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
     4741 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     4801 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     4861 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     4921 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     4981 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     5041 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     5101 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     5161 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     5221 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     5281 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     5341 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     5401 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     5461 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     5521 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     5581 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     5641 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     5701 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     5761 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     5821 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     5881 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     5941 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     6001 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     6061 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     6121 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     6181 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     6241 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     6301 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     6361 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     6421 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6481 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6541 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6601 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6661 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6721 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6781 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6841 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6901 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6961 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     7021 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     7081 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7141 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7201 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7261 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     7321 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     7381 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     7441 agatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     7501 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     7561 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     7621 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     7681 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     7741 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     7801 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     7861 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     7921 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     7981 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     8041 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     8101 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     8161 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     8221 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     8281 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     8341 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     8401 accatccact cgacacaccc gccagcggcc gc
//
