LOCUS       Exported                7649 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133555
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7649)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7649)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7649
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             545..565
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             599..676
                     /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             704..724
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1189..1310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1200..1219)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1254..1273
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1425..1471
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1501..1637
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1682..1753
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1979..2567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2059..2078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3067..4276
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4113..4132
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4280..5308
                     /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    5335..5383
                     /note="synthetic polyadenylation signal"
     CDS             5649..6509
                     /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(5867..5886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6522..6658
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6803..6874
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6879..6925
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6943..6991
                     /note="synthetic polyadenylation signal"
     enhancer        7072..7375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7377..7579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7530..7550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      7648..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggag tctctcgttc ccggctttaa tgaaaaaact catgtccagt tatccttgcc
       61 agtgcttcaa gtaagggatg tgctggtgcg gggtttcgga gacagtgtag aggaagtcct
      121 gtcagaggct agacagcacc tgaaggatgg gacctgtggc ttagtggaag ttgagaaggg
      181 agtcctcccg cagctggaac aaccttatgt gtttataaaa cgcagcgacg cgcgaacagc
      241 acctcacggg catgtaatgg tggagcttgt cgccgagttg gagggcatcc agtacggaag
      301 atcgggtgaa acgctgggag ttctagtgcc ccacgttggc gaaattccgg tggcctaccg
      361 taaggttttg ctcaggaaaa acggaaataa gggggctggt ggccatagct atggggcaga
      421 cctgaagtct ttcgatctgg gcgacgagct gggcaccgac ccatatgagg atttccagga
      481 aaactggaac acaaaacaca gctccggggt aactcgcgag ctaatgcgag aattgaatgg
      541 aggggaaaac ctttactttc agtcgggacg gcgaatcccg gggctgataa atccctggaa
      601 aaggagatgg aagaagaact ttattgcagt gtctgcggct aaccggttca aaaagatcag
      661 ttcaagcggc gccctagatt acgacattcc tacgaccgcc tccgagaatc tctatttcca
      721 gggtgaattt ggcttggcac aacatgacga ggccgtcgat aacaagttta ataaagaaca
      781 gcagaacgct ttctatgaga tccttcactt accaaatctg aacgaagagc aacgcaacgc
      841 tttcattcag agcctgaaag acgatccctc gcagtctgca aatctcctgg ccgaagccaa
      901 gaagctgaat gacgcacagg ctccaaaagt tgataacaag tttaacaagg agcagcagaa
      961 tgctttctac gagatattgc accttcctaa tctgaatgag gaacaaagaa atgcatttat
     1021 ccagtcccta aaagacgatc catcacagag tgccaacctg cttgcggaag ccaaaaagct
     1081 caatgatgcc caggccccga aagtagacgc gaaccatcaa taaggatccg caggcctctg
     1141 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1201 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1261 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1321 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1381 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1441 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     1501 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1561 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1621 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1681 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     1741 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     1801 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     1861 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     1921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     2821 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     2881 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     2941 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3001 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3061 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3121 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3181 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3241 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3301 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3361 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     3421 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     3481 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     3541 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     3601 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     3661 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     3721 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     3781 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     3841 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     3901 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     3961 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4021 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4081 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4141 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4201 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4261 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     4321 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     4381 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4441 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     4501 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     4561 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4621 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     4681 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4741 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4801 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     4861 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4921 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4981 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5041 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     5101 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     5161 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     5221 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     5281 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     5341 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     5401 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     5461 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     5521 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     5581 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     5641 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5761 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5821 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5881 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5941 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6061 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6181 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     6241 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     6301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6481 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     6541 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     6601 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     6661 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     6721 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     6781 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     6841 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     6901 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     6961 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     7021 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7081 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7141 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7201 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7261 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7321 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     7381 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7441 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7501 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7561 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     7621 atccactcga cacacccgcc agcggccgc
//
