LOCUS       Exported                6851 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133233
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6851)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6851
                     /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..322
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    391..512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(402..421)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     456..475
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      627..673
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      703..839
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    884..955
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1181..1769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1261..1280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2269..3478
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3315..3334
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3482..4510
                     /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    4537..4585
                     /note="synthetic polyadenylation signal"
     CDS             4851..5711
                     /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(5069..5088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5724..5860
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6005..6076
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6081..6127
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6145..6193
                     /note="synthetic polyadenylation signal"
     enhancer        6274..6577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6579..6781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6732..6752
                     /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 tcagtcgaaa gaaaccgcgg cagctaagtt
      301 cgagaggcag cacatggaca gttaaggatc cgcaggcctc tgctagcttg actgactgag
      361 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
      421 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
      481 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
      541 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
      601 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
      661 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
      721 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
      781 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
      841 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
      901 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
      961 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     1021 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     1081 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     1141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1621 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1981 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     2041 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     2101 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     2161 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     2221 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
     2281 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
     2341 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     2401 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     2461 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     2521 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     2581 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     2641 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     2701 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     2761 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     2821 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     2881 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
     2941 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
     3001 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     3061 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     3121 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
     3181 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     3241 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     3301 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     3361 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     3421 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
     3481 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     3541 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     3601 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     3661 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga
     3721 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     3781 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat
     3841 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     3901 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     3961 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     4021 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     4081 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4141 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4201 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4261 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4321 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     4381 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga
     4441 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     4501 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga
     4561 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga
     4621 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     4681 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     4741 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     4801 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     4861 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4921 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     4981 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     5041 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5101 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     5161 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5221 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     5281 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5341 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5401 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5461 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5521 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5581 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5641 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5701 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     5761 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     5821 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct
     5881 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     5941 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     6001 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     6061 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     6121 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     6181 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     6241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     6301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     6421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     6481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     6541 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     6601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     6661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     6721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     6781 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     6841 ccagcggccg c
//
