LOCUS Exported 8349 bp ds-DNA circular SYN 10-JAN-2023 DEFINITION Split TEV assays. ACCESSION . VERSION . KEYWORDS pcDNA3_GCGR-V2R-NTEV-TCS-GV-2xHA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8349) TITLE Wehr Split TEV assay plasmids REFERENCE 2 (bases 1 to 8349) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 10, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8349 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 899..923 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2361..2385) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2876..2896 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 2903..2932 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2942..3382 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="DNA binding domain of the GAL4 transcriptional activator" /label=GAL4 DNA binding domain /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS" CDS 3425..3445 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3596..3829 /codon_start=1 /gene="UL48" /product="transcriptional activation domain of herpes simplex virus protein VP16 (Triezenberg et al., 1988; Cousens et al., 1989)" /label=VP16 AD /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG" CDS 3833..3859 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 3863..3889 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(3902..3920) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3903..3920) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3940..3957) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3946..4170 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4216..4644 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4303..4322) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4513..4534 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4653..4673) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4658..4987 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4838..4973 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4900..4919 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5054..5848 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5108..5127) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5718..5737 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6022..6143 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6059..6078) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6113..6132 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6192..6208) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6192..6208) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6205..6227) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6216..6232 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6240..6270) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6285..6306 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6423..6440) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6594..7182) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6674..6693) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7353..8213) /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 7976..7995 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8214..8318) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatcaac 901 aagtttgtac aaaaaagcag gctccaccat gcccccctgc cagccacagc gacccctgct 961 gctgttgctg ctgctgctgg cctgccagcc acaggtcccc tccgctcagg tgatggactt 1021 cctgtttgag aagtggaagc tctacggtga ccagtgtcac cacaacctga gcctgctgcc 1081 ccctcccacg gagctggtgt gcaacagaac cttcgacaag tattcctgct ggccggacac 1141 ccccgccaat accacggcca acatctcctg cccctggtac ctgccttggc accacaaagt 1201 gcaacaccgc ttcgtgttca agagatgcgg gcccgacggt cagtgggtgc gtggaccccg 1261 ggggcagcct tggcgtgatg cctcccagtg ccagatggat ggcgaggaga ttgaggtcca 1321 gaaggaggtg gccaagatgt acagcagctt ccaggtgatg tacacagtgg gctacagcct 1381 gtccctgggg gccctgctcc tcgccttggc catcctgggg ggcctcagca agctgcactg 1441 cacccgcaat gccatccacg cgaatctgtt tgcgtccttc gtgctgaaag ccagctccgt 1501 gctggtcatt gatgggctgc tcaggacccg ctacagccag aaaattggcg acgacctcag 1561 tgtcagcacc tggctcagtg atggagcggt ggctggctgc cgtgtggccg cggtgttcat 1621 gcaatatggc atcgtggcca actactgctg gctgctggtg gagggcctgt acctgcacaa 1681 cctgctgggc ctggccaccc tccccgagag gagcttcttc agcctctacc tgggcatcgg 1741 ctggggtgcc cccatgctgt tcgtcgtccc ctgggcagtg gtcaagtgtc tgttcgagaa 1801 cgtccagtgc tggaccagca atgacaacat gggcttctgg tggatcctgc ggttccccgt 1861 cttcctggcc atcctgatca acttcttcat cttcgtccgc atcgttcagc tgctcgtggc 1921 caagctgcgg gcacggcaga tgcaccacac agactacaag ttccggctgg ccaagtccac 1981 gctgaccctc atccctctgc tgggcgtcca cgaagtggtc ttcgccttcg tgacggacga 2041 gcacgcccag ggcaccctgc gctccgccaa gctcttcttc gacctcttcc tcagctcctt 2101 ccagggcctg ctggtggctg tcctctactg cttcctcaac aaggaggtgc agtcggagct 2161 gcggcggcgt tggcaccgct ggcgcctggg caaagtgcta tgggaggagc ggaacaccag 2221 caaccacagg gcctcatctt cgcccggcca cggccctccc agcaaggagc tgcagtttgg 2281 gaggggtggt ggcagccagg attcatctgc ggagaccccc ttggctggtg gcctccctag 2341 attggctgag agccccttcg acccagcttt cttgtacaaa gtggttgatg gtaccgcccg 2401 gggacgcacc ccacccagcc tgggtcccca agatgagtcc tgcaccaccg ccagctcctc 2461 cctggccaag gacacttcat cggatgtcgg aggtggaggt agcggaggtg gaggtagcgg 2521 agagagcttg tttaagggac cacgtgatta caacccgata tcgagcacca tttgtcattt 2581 gacgaatgaa tctgatgggc acacaacatc gttgtatggt attggatttg gtcccttcat 2641 cattacaaac aagcacttgt ttcgtcgcaa taatggaaca ctgttggtcc aatcactaca 2701 tggtgtattc aaggtcaagg acaccacgac tttgcaacaa cacctcattg atgggaggga 2761 catgatgatt attcgcatgc ctaaggattt cccaccattt cctcaaaagc tgaaatttag 2821 agagccacaa agggaagagc gcatatgtct tgtgacaacc aacttccaag ctcccgagaa 2881 cctgtacttc cagggttcta gagaacagaa gctgatctca gaagaggatc tggcccgggc 2941 catgaagcta ctgtcttcta tcgaacaagc atgcgatatt tgccgactta aaaagctcaa 3001 gtgctccaaa gaaaaaccga agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta 3061 ctctcccaaa accaaaaggt ctccgctgac tagggcacat ctgacagaag tggaatcaag 3121 gctagaaaga ctggaacagc tatttctact gatttttcct cgagaagacc ttgacatgat 3181 tttgaaaatg gattctttac aggatataaa agcattgtta acaggattat ttgtacaaga 3241 taatgtgaat aaagatgccg tcacagatag attggcttca gtggagactg atatgcctct 3301 aacattgaga cagcatagaa taagtgcgac atcatcatcg gaagagagta gtaacaaagg 3361 tcaaagacag ttgactgtat cgccggaatt cccggggatc agcttgataa ttcgattcac 3421 catgcccaag aagaagagga aggtggcgta cagccgcgcg cgtacgaaaa acaattacgg 3481 gtctaccatc gagggcctgc tcgatctccc ggacgacgac gcccccgaag aggcggggct 3541 ggcggctccg cgcctgtcct ttctccccgc gggacacacg cgcagactgt cgacggcccc 3601 cccgaccgat gtcagcctgg gggacgagct ccacttagac ggcgaggacg tcgcgatggc 3661 gcatgccgac gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg 3721 tccgggattt accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga 3781 gtttgagcag atgtttaccg atgcccttgg aattgacgag tacggtgggg gctacccata 3841 cgatgttcct gactatgcgg gctatcccta tgacgtcccg gactatgcat gagggcccta 3901 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt 3961 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc 4021 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt 4081 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca 4141 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct 4201 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 4261 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 4321 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 4381 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 4441 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 4501 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 4561 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 4621 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa 4681 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 4741 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 4801 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 4861 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 4921 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 4981 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg 5041 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 5101 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 5161 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 5221 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 5281 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 5341 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 5401 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 5461 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 5521 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 5581 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 5641 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 5701 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 5761 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 5821 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 5881 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 5941 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 6001 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 6061 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 6121 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 6181 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 6241 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 6301 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6361 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6421 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6481 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6541 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6601 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6661 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6721 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6781 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6841 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6901 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6961 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7021 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7081 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7141 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7201 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 7261 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 7321 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7381 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7441 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7501 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7561 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7621 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7681 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7741 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7801 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7861 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7921 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7981 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8041 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8101 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8161 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 8221 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 8281 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 8341 cctgacgtc //