LOCUS       Exported                7355 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133444
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7355)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7355)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7355
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             305..382
                     /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"
     CDS             410..430
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    895..1016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(906..925)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     960..979
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1131..1177
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1207..1343
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1388..1459
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1685..2273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1765..1784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2773..3982
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3819..3838
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3986..5014
                     /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    5041..5089
                     /note="synthetic polyadenylation signal"
     CDS             5355..6215
                     /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(5573..5592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6228..6364
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6509..6580
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6585..6631
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6649..6697
                     /note="synthetic polyadenylation signal"
     enhancer        6778..7081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7083..7285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7236..7256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc gggacggcga atcccggggc tgataaatcc
      301 ctggaaaagg agatggaaga agaactttat tgcagtgtct gcggctaacc ggttcaaaaa
      361 gatcagttca agcggcgccc tagattacga cattcctacg accgcctccg agaatctcta
      421 tttccagggt gaatttggct tggcacaaca tgacgaggcc gtcgataaca agtttaataa
      481 agaacagcag aacgctttct atgagatcct tcacttacca aatctgaacg aagagcaacg
      541 caacgctttc attcagagcc tgaaagacga tccctcgcag tctgcaaatc tcctggccga
      601 agccaagaag ctgaatgacg cacaggctcc aaaagttgat aacaagttta acaaggagca
      661 gcagaatgct ttctacgaga tattgcacct tcctaatctg aatgaggaac aaagaaatgc
      721 atttatccag tccctaaaag acgatccatc acagagtgcc aacctgcttg cggaagccaa
      781 aaagctcaat gatgcccagg ccccgaaagt agacgcgaac catcaataag gatccgcagg
      841 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      901 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      961 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1021 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     1081 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     1141 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     1201 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     1261 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     1321 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     1381 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     1441 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     1501 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     1561 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     1621 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2101 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     2521 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     2581 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     2641 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     2701 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     2761 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2821 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2881 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2941 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     3001 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     3061 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     3121 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     3181 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     3241 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     3301 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     3361 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     3421 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     3481 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     3541 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     3601 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     3661 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3721 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3781 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3841 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3901 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3961 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     4021 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     4081 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     4141 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     4201 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     4261 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4321 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     4381 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4441 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4501 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4561 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4621 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4681 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4741 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4801 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4861 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4921 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4981 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     5041 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     5101 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     5161 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     5221 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     5281 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     5341 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5401 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5461 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5521 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5581 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5641 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     5701 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5761 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5821 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5881 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5941 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     6001 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6061 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6121 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6181 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     6241 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     6301 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     6361 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     6421 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     6481 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     6541 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     6601 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     6661 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     6721 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     6781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     6841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     7021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     7081 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     7141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     7201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     7261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     7321 cctaccatcc actcgacaca cccgccagcg gccgc
//
