LOCUS       Exported                6852 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252917
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6852
                     /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             272..316
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    385..506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(396..415)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     450..469
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      621..667
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      697..833
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    878..949
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1175..1763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1255..1274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2263..3472
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3309..3328
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3476..4504
                     /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    4531..4579
                     /note="synthetic polyadenylation signal"
     CDS             4845..5705
                     /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(5063..5082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5718..5854
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6006..6077
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6082..6128
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6146..6194
                     /note="synthetic polyadenylation signal"
     enhancer        6275..6578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6580..6782
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6733..6753
                     /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 gaaagaaacc gcggcagcta agttcgagag
      301 gcagcacatg gacagttaag gatccgcagg cctctgctag cttgactgac tgagatacag
      361 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
      421 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      481 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      541 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
      601 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
      661 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
      721 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
      781 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
      841 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
      901 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
      961 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     1021 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     1081 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     1141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1981 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     2041 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     2101 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     2161 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2221 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2281 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2341 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2401 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2461 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2521 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2581 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2641 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     2701 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     2761 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     2821 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     2881 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     2941 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     3001 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     3061 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     3121 ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg
     3181 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3241 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3301 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3361 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     3421 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     3481 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3541 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3601 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3661 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     3721 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3781 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     3841 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3901 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3961 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4021 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4081 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4141 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4201 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4261 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4321 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4381 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     4441 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4501 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     4561 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     4621 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     4681 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     4741 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     4801 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     4861 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4921 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     4981 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     5041 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5101 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5161 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5221 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5281 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5341 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5401 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     5461 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5521 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5581 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5641 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5701 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     5761 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     5821 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     5881 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     5941 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     6001 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     6061 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     6121 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     6181 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     6241 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6301 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6361 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6421 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6481 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6541 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     6601 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6661 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6721 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6781 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     6841 gccagcggcc gc
//
