LOCUS Exported 8031 bp ds-DNA circular SYN 10-JAN-2023 DEFINITION Split TEV assays. ACCESSION . VERSION . KEYWORDS pcDNA3_AVPR2-V2R-NTEV-TCS-GV-2xHA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8031) TITLE Wehr Split TEV assay plasmids REFERENCE 2 (bases 1 to 8031) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 10, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8031 /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(2043..2067) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2558..2578 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 2585..2614 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2624..3064 /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 3107..3127 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3278..3511 /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 3515..3541 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 3545..3571 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(3584..3602) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3585..3602) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3622..3639) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3628..3852 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3898..4326 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3985..4004) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4195..4216 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4335..4355) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4340..4669 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4520..4655 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4582..4601 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4736..5530 /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(4790..4809) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5400..5419 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5704..5825 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5741..5760) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5795..5814 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5874..5890) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5874..5890) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5887..5909) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5898..5914 /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(5922..5952) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5967..5988 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6105..6122) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6276..6864) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6356..6375) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7035..7895) /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 7658..7677 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7896..8000) /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 gctctaccat gctcatggcg tccaccactt ccgctgtgcc 961 tgggcatccc tctctgccca gcctgcccag caacagcagc caggagaggc cactggacac 1021 ccgggacccg ctgctagccc gggcggagct ggcgctgctc tccatagtct ttgtggctgt 1081 ggccctgagc aatggcctgg tgctggcggc cctagctcgg cggggccggc ggggccactg 1141 ggcacccata cacgtcttca ttggccactt gtgcctggcc gacctggccg tggctctgtt 1201 ccaagtgctg ccccagctgg cctggaaggc caccgaccgc ttccgtgggc cagatgccct 1261 gtgtcgggcc gtgaagtatc tgcagatggt gggcatgtat gcctcctcct acatgatcct 1321 ggccatgacg ctggaccgcc accgtgccat ctgccgtccc atgctggcgt accggcatgg 1381 aagtggggct cactggaacc ggccggtgct agtggcttgg gccttctcgc tccttctcag 1441 cctgccccag ctcttcatct tcgcccagcg caacgtggaa ggtggcagcg gggtcactga 1501 ctgctgggcc tgctttgcgg agccctgggg ccgtcgcacc tatgtcacct ggattgccct 1561 gatggtgttc gtggcaccta ccctgggtat cgccgcctgc caggtgctca tcttccggga 1621 gattcatgcc agtctggtgc cagggccatc agagaggcct ggggggcgcc gcaggggacg 1681 ccggacaggc agccccggtg agggagccca cgtgtcagca gctgtggcca agactgtgag 1741 gatgacgcta gtgattgtgg tcgtctatgt gctgtgctgg gcacccttct tcctggtgca 1801 gctgtgggcc gcgtgggacc cggaggcacc tctggaaggg gcgccctttg tgctactcat 1861 gttgctggcc agcctcaaca gctgcaccaa cccctggatc tatgcatctt tcagcagcag 1921 cgtgtcctca gagctgcgaa gcttgctctg ctgtgcccgg ggacgcaccc cacccagcct 1981 gggtccccaa gatgagtcct gcaccaccgc cagctcctcc ctggccaagg acacttcatc 2041 ggacccagct ttcttgtaca aagtggttga tggtaccgcc cggggacgca ccccacccag 2101 cctgggtccc caagatgagt cctgcaccac cgccagctcc tccctggcca aggacacttc 2161 atcggatgtc ggaggtggag gtagcggagg tggaggtagc ggagagagct tgtttaaggg 2221 accacgtgat tacaacccga tatcgagcac catttgtcat ttgacgaatg aatctgatgg 2281 gcacacaaca tcgttgtatg gtattggatt tggtcccttc atcattacaa acaagcactt 2341 gtttcgtcgc aataatggaa cactgttggt ccaatcacta catggtgtat tcaaggtcaa 2401 ggacaccacg actttgcaac aacacctcat tgatgggagg gacatgatga ttattcgcat 2461 gcctaaggat ttcccaccat ttcctcaaaa gctgaaattt agagagccac aaagggaaga 2521 gcgcatatgt cttgtgacaa ccaacttcca agctcccgag aacctgtact tccagggttc 2581 tagagaacag aagctgatct cagaagagga tctggcccgg gccatgaagc tactgtcttc 2641 tatcgaacaa gcatgcgata tttgccgact taaaaagctc aagtgctcca aagaaaaacc 2701 gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc tactctccca aaaccaaaag 2761 gtctccgctg actagggcac atctgacaga agtggaatca aggctagaaa gactggaaca 2821 gctatttcta ctgatttttc ctcgagaaga ccttgacatg attttgaaaa tggattcttt 2881 acaggatata aaagcattgt taacaggatt atttgtacaa gataatgtga ataaagatgc 2941 cgtcacagat agattggctt cagtggagac tgatatgcct ctaacattga gacagcatag 3001 aataagtgcg acatcatcat cggaagagag tagtaacaaa ggtcaaagac agttgactgt 3061 atcgccggaa ttcccgggga tcagcttgat aattcgattc accatgccca agaagaagag 3121 gaaggtggcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca tcgagggcct 3181 gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc cgcgcctgtc 3241 ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg atgtcagcct 3301 gggggacgag ctccacttag acggcgagga cgtcgcgatg gcgcatgccg acgcgctaga 3361 cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat ttacccccca 3421 cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc agatgtttac 3481 cgatgccctt ggaattgacg agtacggtgg gggctaccca tacgatgttc ctgactatgc 3541 gggctatccc tatgacgtcc cggactatgc atgagggccc tattctatag tgtcacctaa 3601 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 3661 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 3721 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 3781 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 3841 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 3901 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 3961 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 4021 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 4081 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 4141 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 4201 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 4261 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 4321 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 4381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 4441 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 4501 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 4561 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 4621 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 4681 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat 4741 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 4801 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 4861 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 4921 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 4981 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 5041 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 5101 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 5161 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 5221 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 5281 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 5341 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 5401 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 5461 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 5521 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 5581 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 5641 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 5701 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 5761 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5821 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 5881 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 5941 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 6001 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 6061 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 6121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 6181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 6241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 6301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 6361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 6421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 6481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 6541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 6601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 6661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 6721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 6781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 6961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 7021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 7081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 7141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 7201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 7261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 7321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 7381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 7441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 7501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 7561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 7621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 7681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 7801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //