LOCUS Exported 10331 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses HA tagged Citrine or FLAG tagged Cerulean upon binding of TALE1 to VAS1 or TALE4 to VAS4. ACCESSION . VERSION . KEYWORDS pJFRC81-3xVAS1-Syn21-Citrine-HA-P10-3xVAS4-Syn21-FLAG-Cerulean... SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10331) AUTHORS Toegel M, Azzam G, Lee EY, Knapp DJHF, Tan Y, Fa M, Fulga TA TITLE A multiplexable TALE-based binary expression system for in vivo cellular interaction studies. JOURNAL Nat Commun. 2017 Nov 21;8(1):1663. doi: 10.1038/s41467-017-01592-3. PUBMED 29162808 REFERENCE 2 (bases 1 to 10331) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10331 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 31..135 /gene="bla" /label=AmpR promoter CDS 136..996 /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(354..373) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1167..1755 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1656..1675 /label=pBR322ori-F /note="pBR322 origin, forward primer" protein_bind 1842..1911 /label=attB /bound_moiety="phage phi-C31 integrase" /note="attB site for the phi-C31 integrase (Groth et al., 2000)" primer_bind complement(2136..2154) /label=pCasper-F /note="Drosophila mini-white gene, reverse primer" CDS join(2606..2677,3079..3352,3427..4081,4143..4458,4662..4793, 4864..5478) /codon_start=1 /gene="white" /product="Drosophila white gene eye color pigment" /label=mini-white /note="This is a modified version of the white gene lacking part of the first intron." /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE" promoter 6569..6807 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" CDS 6860..7576 /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 7583..7609 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(9826..9849) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(10277..10295) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 gactcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 61 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 1021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 1081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 1141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 1201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 1261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 1321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 1381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 1441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 1501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 1561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 1621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 1681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 1741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 1801 ccttttgctc acatgttacc gtcgacgatg taggtcacgg tctcgaagcc gcggtgcggg 1861 tgccagggcg tgcccttggg ctccccgggc gcgtactcca cctcacccat ctggtccatc 1921 atgatgaacg ggtcgaggtg gcggtagttg atcccggcga acgcgcggcg caccgggaag 1981 ccctcgccct cgaaaccgct gggcgcggtg gtcacggtga gcacgggacg tgcgacggcg 2041 tcggcgggtg cggatacgcg gggcagcgtc agcgggttct cgacggtcac ggcgggcatg 2101 tcgacaagcc gaacatatgg gcgcgcctag tatgtatgta agttaataaa acccattttt 2161 gcggaaagta gataaaaaaa acattttttt tttttactgc actggatatc attgaactta 2221 tctgatcagt tttaaattta cttcgatcca agggtatttg atgtaccagg ttctttcgat 2281 tacctctcac tcaaaatgac attccactca aagtcagcgc tgtttgcctc cttctctgtc 2341 cacagaaata tcgccgtctc tttcgccgct gcgtccgcta tctctttcgc caccgtttgt 2401 agcgttacgt agcgtcaatg tccgccttca gttgcatttt gtcagcggtt tcgtgacgaa 2461 gctccaagcg gtttacgcca tcaattaaac acaaagtgct gtgccaaaac tcctctcgct 2521 tcttattttt gtttgttttt tgagtgattg gggtggtgat tggttttggg tgggtaagca 2581 ggggaaagtg tgaaaaatcc cggcaatggg ccaagaggat caggagctat taattcgcgg 2641 aggcagcaaa cacccatctg ccgagcatct gaacaatgtg agtagtacat gtgcatacat 2701 cttaagttca cttgatctat aggaactgcg attgcaacat caaattgtct gcggcgtgag 2761 aactgcgacc cacaaaaatc ccaaaccgca attgcacaaa caaatagtga cacgaaacag 2821 attattctgg tagctgttct cgctatataa gacaattttt gagatcatat catgatcaag 2881 acatctaaag gcattcattt tcgactatat tcttttttac aaaaaatata acaaccagat 2941 attttaagct gatcctagat gcacaaaaaa taaataaaag tataaaccta cttcgtagga 3001 tacttcgggg tactttttgt tcggggttag atgagcataa cgcttgtagt tgatatttga 3061 gatcccctat cattgcaggg tgacagcgga gcggcttcgc agagctgcat taaccagggc 3121 ttcgggcagg ccaaaaacta cggcacgctc cggccaccca gtccgccgga ggactccggt 3181 tcagggagcg gccaactagc cgagaacctc acctatgcct ggcacaatat ggacatcttt 3241 ggggcggtca atcagccggg ctccggatgg cggcagctgg tcaaccggac acgcggacta 3301 ttctgcaacg agcgacacat accggcgccc aggaaacatt tgctcaagaa cggtgagttt 3361 ctattcgcag tcggctgatc tgtgtgaaat cttaataaag ggtccaatta ccaatttgaa 3421 actcagtttg cggcgtggcc tatccgggcg aacttttggc cgtgatgggc agttccggtg 3481 ccggaaagac gaccctgctg aatgcccttg cctttcgatc gccgcagggc atccaagtat 3541 cgccatccgg gatgcgactg ctcaatggcc aacctgtgga cgccaaggag atgcaggcca 3601 ggtgcgccta tgtccagcag gatgacctct ttatcggctc cctaacggcc agggaacacc 3661 tgattttcca agccatggtg cggatgccac gacatctgac ctatcggcag cgagtggccc 3721 gcgtggatca ggtgatccag gagctttcgc tcagcaaatg tcagcacacg atcatcggtg 3781 tgcccggcag ggtgaaaggt ctgtccggcg gagaaaggaa gcgtctggca ttcgcctccg 3841 aggctctaac cgatccgccg cttctgatct gcgatgagcc cacctccgga ctggactcct 3901 ttaccgccca cagcgtcgtc caggtgctga agaagctgtc gcagaagggc aagaccgtca 3961 tcctgaccat tcatcagccg tcttccgagc tgtttgagct ctttgacaag atccttctga 4021 tggccgaggg cagggtagct ttcttgggca ctcccagcga agccgtcgac ttcttttcct 4081 agtgagttcg atgtgtttat taagggtatc tagtattaca taacatctca actcctatcc 4141 agcgtgggtg cccagtgtcc taccaactac aatccggcgg acttttacgt acaggtgttg 4201 gccgttgtgc ccggacggga gatcgagtcc cgtgatcgga tcgccaagat atgcgacaat 4261 tttgccatta gcaaagtagc ccgggatatg gagcagttgt tggccaccaa aaatctggag 4321 aagccactgg agcagccgga gaatgggtac acctacaagg ccacctggtt catgcagttc 4381 cgggcggtcc tgtggcgatc ctggctgtcg gtgctcaagg aaccactcct cgtaaaagtg 4441 cgacttattc agacaacggt gagtggttcc agtggaaaca aatgatataa cgcttacaat 4501 tcttggaaac aaattcgcta gattttagat agaattgcct gattccacac ccttcttagt 4561 ttttttcaat gagatgtata gtttatagtt ttgcagaaga taaataaatt tcatttaact 4621 cgcgaatatt aatgagatgc gagtaacatt ttaatttgca gatggttgcc atcttgattg 4681 gcctcatctt tttgggccaa caactcacgc aagtgggtgt gatgaatatc aacggagcca 4741 tcttcctctt cctgaccaac atgacctttc aaaacgtctt tgccacgata aatgtaagtc 4801 atgtttagaa tacatttgca tttcaataat ttactaactt tctaatgaat cgattcgatt 4861 taggtgttca cctcagagct gccagttttt atgagggagg cccgaagtcg actttatcgc 4921 tgtgacacat actttctggg caaaacgatt gccgaattgc cgctttttct cacagtgcca 4981 ctggtcttca cggcgattgc ctatccgatg atcggactgc gggccggagt gctgcacttc 5041 ttcaactgcc tggcgctggt cactctggtg gccaatgtgt caacgtcctt cggatatcta 5101 atatcctgcg ccagctcctc gacctcgatg gcgctgtctg tgggtccgcc ggttatcata 5161 ccattcctgc tctttggcgg cttcttcttg aactcgggct cggtgccagt atacctcaaa 5221 tggttgtcgt acctctcatg gttccgttac gccaacgagg gtctgctgat taaccaatgg 5281 gcggacgtgg agccgggcga aattagctgc acatcgtcga acaccacgtg ccccagttcg 5341 ggcaaggtca tcctggagac gcttaacttc tccgccgccg atctgccgct ggactacgtg 5401 ggtctggcca ttctcatcgt gagcttccgg gtgctcgcat atctggctct aagacttcgg 5461 gcccgacgca aggagtagcc gacatatatc cgaaataact gcttgttttt tttttttacc 5521 attattacca tcgtgtttac tgtttattgc cccctcaaaa agctaatgta attatatttg 5581 tgccaataaa aacaagatat gacctataga atacaagtat ttccccttcg aacatcccca 5641 caagtagact ttggatttgt cttctaacca aaagacttac acacctgcat accttacatc 5701 aaaaactcgt ttatcgctac ataaaacacc gggatatatt ttttatatac atacttttca 5761 aatcgcgcgc cctcttcata attcacctcc accacaccac gtttcgtagt tgctctttcg 5821 ctgtctccca cccgctctcc gcaacacatt caccttttgt tcgacgacct tggagcgact 5881 gtcgttagtt ccgcgcgatt cggttcgctc aaatggttcc gagtggttca tttcgtctca 5941 atagaaatta gtaataaata tttgtatgta caatttattt gctccaatat atttgtatat 6001 atttccctca cagctatatt tattctaatt taatattatg actttttaag gtaatttttt 6061 gtgacctgtt cggagtgatt agcgttacaa tttgaactga aagtgacatc cagtgtttgt 6121 tccttgtgta gatgcatctc aaaaaaatgg tgggcataat agtgttgttt atatatatca 6181 aaaataacaa ctataataat aagaatacat ttaatttaga aaatgcttgg atttcactgg 6241 aactagggcg cgcctccgga acataatggt gcagggcgct gacttccgcg tttccagact 6301 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca 6361 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca 6421 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca 6481 gggccgcaag cttagtcgac cccgcgggta gttagtcgac cccgcgggta gttagtcgac 6541 cccgcgggta gtctagcgac gtcgagcgcc ggagtataaa tagaggcgct tcgtctacgg 6601 agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat 6661 aaacaagcgc agctgaacaa gctaaacaat ctgcagtaaa gtgcaagtta aagtgaatca 6721 attaaaagta accagcaacc aagtaaatca actgcaacta ctgaaatctg ccaagaagta 6781 attattgaat acaagaagag aactctgaat agatctgcgg ccgcggctcg agggtaccaa 6841 cttaaaaaaa aaaatcaaaa tggtcagcaa gggcgaggag ctgttcaccg gggtggtgcc 6901 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 6961 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 7021 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt gcttcgcccg 7081 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 7141 ccaggagcgc accatcttct tcaaagacga cggcaactac aagacccgcg ccgaggtgaa 7201 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 7261 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 7321 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 7381 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 7441 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga 7501 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 7561 ggacgagctg tacaagggtg gatacccata cgatgttcca gattacgctt aatagtctag 7621 aatgaatcgt ttttaaaata acaaatcaat tgttttataa tattcgtacg attctttgat 7681 tatgtaataa aatgtgatca ttaggaagat tacgaaaaat ataaaaaata tgagttctgt 7741 gtgtataaca aatgctgtaa acgccacaat tgtgtttgtt gcaaataaac ccatgattat 7801 ttgattaaaa ttgttgtttt ctttgttcat atacaatagt gtgttttgcc taaacgtgta 7861 ctgcataaac tccatgcgag tgtatagcga gctagtggct aacgcttgcc ccaccaaagt 7921 agattcgtca aaatcctcaa tttcatcacc ctcctccaag tttaacattt ggccgtcgga 7981 attaacttct aaagatgcca cataatstaa taaatgaaat agagattcaa acgtggcgtc 8041 atcgtccgtt tcgaccattt ccgaaaagaa ctcgggcata aactctatga tttctctgga 8101 cgtggtgttg tcgaaactct caaagtacgc agtcaggaac gtgcgcgaca tgtcgtcggg 8161 aaactcgcgc ggaaacatgt tgttgtaacc gaacgggtcc catagcgcca aaaccaaatc 8221 tgccagcgtc aatagaatga gcacgatgcc gacaatggag ctggcttgga tagcgattcg 8281 agttaacgaa ttcttgaaga cgaaagggga attcagaagt tcaaatgggc tttcatattt 8341 acttttttct ctgagtgggt aaggtcattg tggggtctta agggctaaag ggtagtgatg 8401 atgaaggcat ttgatgccat tgttcaataa cttaccgctg gggttaactc gaatcgctat 8461 ccaagccagc tccattgtcg gcatcgtgct cattctattg acgctggcag atttggtttt 8521 ggcgctatgg gacccgttcg gttacaacaa catgtttccg cgcgagtttc ccgacgacat 8581 gtcgcgcacg ttcctgactg cgtactttga gagtttcgac aacaccacgt ccagagaaat 8641 catagagttt atgcccgagt tcttttcgga aatggtcgaa acggacgatg acgccacgtt 8701 tgaatctcta tttcatttat tagattatgt ggcatcttta gaagttaatt ccgacggcca 8761 aatgttaaac ttggaggagg gtgatgaaat tgaggatttt gacgaatcta ctttggtggg 8821 gcaagcgtta gccactagct cgctatacac tcgcatggag tttatgcagt acacgtttag 8881 gcaaaacaca ctattgtcta tgaacaaaga aaacaacaat tttaatcaaa taatcatggg 8941 tttatttgca acaaacacaa ttgtggcgtt tacagcattt gttatacaca cagaactcat 9001 attttttata tttttcgtaa tcttcctaat gatcacattt tattacataa tcaaagaatc 9061 gtacgaatat tataaaacaa ttgatttgtt attttaaaaa cgattcattt tacttgtaca 9121 actctccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc 9181 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg 9241 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac 9301 gctgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt 9361 gtcggcggtg atatagacgt tgtcgctgat ggcgttgtac tccagcttgt gccccaggat 9421 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc 9481 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tctttgaaga agatggtgcg 9541 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc 9601 ggggtagcgg gcgaagcact gcacgcccca ggtcagggtg gtcacgaggg tgggccaggg 9661 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc 9721 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac 9781 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccttgt cgtcgtcgtc 9841 cttgtagtca ccagccattt tgattttttt ttttatgttg gtaccctcga gccgcggccg 9901 cttgtattca ataattactt cttggcatat ttcagtagtt gcagttgatt tacttggttg 9961 ctggttactt ttaattgatt cactttaact tgcactttac tgcagattgt ttagcttgtt 10021 cagctgcgct tgtttatttg cttacctttc gcttagcgac gtgttcactt tgcttgtttg 10081 aattgaattg tcgctccgta gacgaagcgc ctctatttat actccggcgg acgtcgctag 10141 aggcaagccc gggtactcta aggcaagccc gggtactcta aggcaagccc gggtactcta 10201 gctagcaagc ttgcggccct ggccacgggt gcgcatgatc gtgctcctgt cgttgagaat 10261 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata 10321 atggtttctt a //