LOCUS       Exported                5010 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133426
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5010)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5010)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5010
                     /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    879..1000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(890..909)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     944..963
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1115..1161
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1191..1327
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1372..1443
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1669..2257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1749..1768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3003..3863
                     /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(3221..3240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3876..4012
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4164..4235
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      4240..4286
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4304..4352
                     /note="synthetic polyadenylation signal"
     enhancer        4433..4736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4738..4940
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4891..4911
                     /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 ctagcttgac
      841 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      901 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      961 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1021 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1081 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     1141 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     1201 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     1261 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     1321 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     1381 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1441 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1501 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1561 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2101 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2341 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2401 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2461 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2521 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2581 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2641 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2701 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     2761 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2821 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2881 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2941 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     3001 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3061 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3121 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3181 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3241 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3301 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3361 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3421 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3481 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3541 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     3601 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     3661 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3721 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3781 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3841 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3901 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3961 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     4021 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     4081 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     4141 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     4201 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     4261 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     4321 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     4381 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4441 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4501 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4561 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4621 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4681 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4741 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4801 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4861 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4921 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4981 catccactcg acacacccgc cagcggccgc
//
