LOCUS       Exported                6884 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133140
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6884)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6884
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             278..355
                     /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    424..545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(435..454)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     489..508
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      660..706
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      736..872
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    917..988
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1214..1802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1294..1313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2302..3511
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3348..3367
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3515..4543
                     /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    4570..4618
                     /note="synthetic polyadenylation signal"
     CDS             4884..5744
                     /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(5102..5121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5757..5893
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6038..6109
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6114..6160
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6178..6226
                     /note="synthetic polyadenylation signal"
     enhancer        6307..6610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6612..6814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6765..6785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaagaaa acctttactt tcagtcgaag aggcgatgga aaaagaattt
      301 tatcgcggtg agtgcagcca acagattcaa aaagatatcc tcgtctggtg ccctctaagg
      361 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
      421 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      481 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      541 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      601 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
      661 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
      721 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
      781 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
      841 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
      901 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
      961 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     1021 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     1081 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     1141 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1621 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1921 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1981 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     2041 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     2101 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     2161 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     2221 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     2281 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     2341 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     2401 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2461 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     2521 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     2581 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     2641 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     2701 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     2761 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     2821 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     2881 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     2941 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     3001 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     3061 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     3121 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     3181 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     3241 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     3301 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     3361 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     3421 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     3481 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     3541 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     3601 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     3661 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     3721 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     3781 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     3841 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     3901 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     3961 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     4021 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     4081 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     4141 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4201 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     4261 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     4321 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     4381 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     4441 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     4501 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     4561 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     4621 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     4681 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     4741 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     4801 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     4861 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     4921 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4981 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     5041 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     5101 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5161 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     5221 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5281 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     5341 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5401 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     5461 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     5521 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5581 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5641 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5701 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     5761 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     5821 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     5881 aatttatcct taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     5941 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     6001 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     6061 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     6121 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     6181 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     6241 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     6361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     6421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     6481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     6541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6601 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     6661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     6721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     6781 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     6841 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
