LOCUS Exported 8112 bp ds-DNA circular SYN 10-JAN-2023 DEFINITION Split TEV assays. ACCESSION . VERSION . KEYWORDS pcDNA3_SP-AVPR2-V2R-NTEV-TCS-GV-2xHA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8112) TITLE Wehr Split TEV assay plasmids REFERENCE 2 (bases 1 to 8112) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 10, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8112 /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(2124..2148) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2639..2659 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 2666..2695 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2705..3145 /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 3188..3208 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3359..3592 /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 3596..3622 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 3626..3652 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(3665..3683) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3666..3683) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3703..3720) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3709..3933 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3979..4407 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4066..4085) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4276..4297 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4416..4436) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4421..4750 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4601..4736 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4663..4682 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4817..5611 /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(4871..4890) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5481..5500 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5785..5906 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5822..5841) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5876..5895 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5955..5971) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5955..5971) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5968..5990) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5979..5995 /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(6003..6033) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6048..6069 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6186..6203) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6357..6945) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6437..6456) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7116..7976) /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 7739..7758 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7977..8081) /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 gaagacgatc atcgccctga gctacatctt 961 ctgcctggta ttcgccgact acaaggacga tgatgacgcc agcatcgata tgctcatggc 1021 gtccaccact tccgctgtgc ctgggcatcc ctctctgccc agcctgccca gcaacagcag 1081 ccaggagagg ccactggaca cccgggaccc gctgctagcc cgggcagagc tggcgctgct 1141 ctccatagtc tttgtggctg tggccctgag caatggcctg gtgctggcgg ccctagctcg 1201 gcggggccgg cggggccact gggcacccat acacgtcttc attggccact tgtgcctggc 1261 cgacctggcc gtggctctgt tccaagtgct gccccagctg gcctggaagg ccaccgaccg 1321 cttccgtggg ccagatgccc tgtgtcgggc cgtgaagtat ctgcagatgg tgggcatgta 1381 tgcctcctcc tacatgatcc tggccatgac gctggaccgc caccgtgcca tctgccgtcc 1441 catgctggcg taccggcatg gaagtggggc tcactggaac cggccggtgc tagtggcttg 1501 ggccttctcg ctccttctca gcctgcccca gctcttcatc ttcgcccagc gcaacgtgga 1561 aggtggcagc ggggtcactg actgctgggc ctgctttgcg gagccctggg gccgtcgcac 1621 ctatgtcacc tggattgccc tgatggtgtt cgtggcacct accctgggta tcgccgcctg 1681 ccaggtgctc atcttccggg agattcatgc cagtctggtg ccagggccat cagagaggcc 1741 tggggggcgc cgcaggggac gccggacagg cagccccggt gagggagccc acgtgtcagc 1801 agctgtggcc aagactgtga ggatgacgct agtgattgtg gtcgtctatg tgctgtgctg 1861 ggcacccttc ttcctggtgc agctgtgggc cgcgtgggac ccggaggcac ctctggaagg 1921 ggcgcccttt gtgctactca tgttgctggc cagcctcaac agctgcacca acccctggat 1981 ctatgcatct ttcagcagca gcgtgtcctc agagctgcga agcttgctct gctgtgcccg 2041 gggacgcacc ccacccagcc tgggtcccca agatgagtcc tgcaccaccg ccagctcctc 2101 cctggccaag gacacttcat cggacccagc tttcttgtac aaagtggttg atggtaccgc 2161 ccggggacgc accccaccca gcctgggtcc ccaagatgag tcctgcacca ccgccagctc 2221 ctccctggcc aaggacactt catcggatgt cggaggtgga ggtagcggag gtggaggtag 2281 cggagagagc ttgtttaagg gaccacgtga ttacaacccg atatcgagca ccatttgtca 2341 tttgacgaat gaatctgatg ggcacacaac atcgttgtat ggtattggat ttggtccctt 2401 catcattaca aacaagcact tgtttcgtcg caataatgga acactgttgg tccaatcact 2461 acatggtgta ttcaaggtca aggacaccac gactttgcaa caacacctca ttgatgggag 2521 ggacatgatg attattcgca tgcctaagga tttcccacca tttcctcaaa agctgaaatt 2581 tagagagcca caaagggaag agcgcatatg tcttgtgaca accaacttcc aagctcccga 2641 gaacctgtac ttccagggtt ctagagaaca gaagctgatc tcagaagagg atctggcccg 2701 ggccatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct 2761 caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg 2821 ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc 2881 aaggctagaa agactggaac agctatttct actgattttt cctcgagaag accttgacat 2941 gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca 3001 agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc 3061 tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa 3121 aggtcaaaga cagttgactg tatcgccgga attcccgggg atcagcttga taattcgatt 3181 caccatgccc aagaagaaga ggaaggtggc gtacagccgc gcgcgtacga aaaacaatta 3241 cgggtctacc atcgagggcc tgctcgatct cccggacgac gacgcccccg aagaggcggg 3301 gctggcggct ccgcgcctgt cctttctccc cgcgggacac acgcgcagac tgtcgacggc 3361 ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg acgtcgcgat 3421 ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc 3481 gggtccggga tttacccccc acgactccgc cccctacggc gctctggata tggccgactt 3541 cgagtttgag cagatgttta ccgatgccct tggaattgac gagtacggtg ggggctaccc 3601 atacgatgtt cctgactatg cgggctatcc ctatgacgtc ccggactatg catgagggcc 3661 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta 3721 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca 3781 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc 3841 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata 3901 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg 3961 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 4021 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 4081 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 4141 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 4201 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 4261 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 4321 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 4381 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg 4441 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 4501 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 4561 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 4621 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 4681 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 4741 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg 4801 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 4861 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 4921 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 4981 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 5041 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 5101 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 5161 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 5221 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 5281 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 5341 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 5401 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 5461 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 5521 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 5581 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 5641 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 5701 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 5761 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 5821 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 5881 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 5941 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 6001 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 6061 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 6121 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 6181 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 6241 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 6301 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 6361 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 6421 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 6481 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 6541 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 6601 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 6661 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 6721 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 6781 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 6841 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 6901 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 6961 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 7021 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 7081 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 7141 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 7201 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 7261 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 7321 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 7381 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 7441 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 7501 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 7561 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 7621 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 7681 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 7741 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 7801 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 7861 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 7921 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 7981 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 8041 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 8101 ccacctgacg tc //