LOCUS       Exported                7368 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252946
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7368
                     /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             311..388
                     /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             416..436
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    901..1022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(912..931)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     966..985
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1137..1183
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1213..1349
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1394..1465
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1691..2279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1771..1790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2779..3988
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3825..3844
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3992..5020
                     /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    5047..5095
                     /note="synthetic polyadenylation signal"
     CDS             5361..6221
                     /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(5579..5598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6234..6370
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6522..6593
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6598..6644
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6662..6710
                     /note="synthetic polyadenylation signal"
     enhancer        6791..7094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7096..7298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7249..7269
                     /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 tcagtcggga cggcgaatcc cggggctgat
      301 aaatccctgg aaaaggagat ggaagaagaa ctttattgca gtgtctgcgg ctaaccggtt
      361 caaaaagatc agttcaagcg gcgccctaga ttacgacatt cctacgaccg cctccgagaa
      421 tctctatttc cagggtgaat ttggcttggc acaacatgac gaggccgtcg ataacaagtt
      481 taataaagaa cagcagaacg ctttctatga gatccttcac ttaccaaatc tgaacgaaga
      541 gcaacgcaac gctttcattc agagcctgaa agacgatccc tcgcagtctg caaatctcct
      601 ggccgaagcc aagaagctga atgacgcaca ggctccaaaa gttgataaca agtttaacaa
      661 ggagcagcag aatgctttct acgagatatt gcaccttcct aatctgaatg aggaacaaag
      721 aaatgcattt atccagtccc taaaagacga tccatcacag agtgccaacc tgcttgcgga
      781 agccaaaaag ctcaatgatg cccaggcccc gaaagtagac gcgaaccatc aataaggatc
      841 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
      901 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      961 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1021 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1081 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1141 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     1201 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1261 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1321 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     1381 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1441 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     1501 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     1561 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1621 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1681 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1741 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1801 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1861 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1921 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1981 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2041 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2101 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2161 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2221 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2281 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2341 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2401 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2461 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     2521 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     2581 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     2641 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     2701 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2761 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2821 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     2881 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     2941 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3001 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3061 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3121 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     3181 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     3241 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     3301 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     3361 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3421 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     3481 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     3541 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3601 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3661 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3721 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3781 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3841 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     3901 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     3961 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     4021 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4081 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4141 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4201 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     4261 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4321 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     4381 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4441 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4501 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4561 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4621 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4681 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4741 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4801 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4861 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4921 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4981 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     5041 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     5101 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     5161 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     5221 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     5281 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     5341 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     5401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5461 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5521 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     5581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5641 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     5701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5761 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     5941 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     6001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6181 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     6241 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     6301 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     6361 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     6421 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     6481 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     6541 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     6601 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     6661 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     6721 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     6781 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6841 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6901 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6961 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     7021 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     7081 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     7141 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     7201 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     7261 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     7321 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
