LOCUS Exported 8332 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses FLAG tagged Cerulean upon binding of TALE4 to VAS4. ACCESSION . VERSION . KEYWORDS pJFRC81-3xVAS4-Syn21-FLAG-Cerulean-P10 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8332) 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 8332) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8332 /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" CDS 6851..6874 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(6917..6938) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7178..7197) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7525..7546 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(8278..8296) /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 cttgctagct agagtacccg ggcttgcctt agagtacccg ggcttgcctt 6541 agagtacccg ggcttgcctc tagcgacgtc cgccggagta taaatagagg cgcttcgtct 6601 acggagcgac aattcaattc aaacaagcaa agtgaacacg tcgctaagcg aaaggtaagc 6661 aaataaacaa gcgcagctga acaagctaaa caatctgcag taaagtgcaa gttaaagtga 6721 atcaattaaa agtaaccagc aaccaagtaa atcaactgca actactgaaa tatgccaaga 6781 agtaattatt gaatacaagc ggccgcggct cgagggtacc aacataaaaa aaaaaatcaa 6841 aatggctggt gactacaagg acgacgacga caaggtgagc aagggcgagg agctgttcac 6901 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt 6961 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac 7021 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca 7081 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc 7141 cgaaggctac gtccaggagc gcaccatctt cttcaaagac gacggcaact acaagacccg 7201 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga 7261 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacgcca tcagcgacaa 7321 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca 7381 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg 7441 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa 7501 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat 7561 cactctcggc atggagagtt gtacaagtaa aatgaatcgt ttttaaaata acaaatcaat 7621 tgttttataa tattcgtacg attctttgat tatgtaataa aatgtgatca ttaggaagat 7681 tacgaaaaat ataaaaaata tgagttctgt gtgtataaca aatgctgtaa acgccacaat 7741 tgtgtttgtt gcaaataaac ccatgattat ttgattaaaa ttgttgtttt ctttgttcat 7801 agacaatagt gtgttttgcc taaacgtgta ctgcataaac tccatgcgag tgtatagcga 7861 gctagtggct aacgcttgcc ccaccaaagt agattcgtca aaatcctcaa tttcatcacc 7921 ctcctccaag tttaacattt ggccgtcgga attaacttct aaagatgcca cataatgtaa 7981 taaatgaaat agagattcaa acgtggcgtc atcgtccgtt tcgaccattt ccgaaaagaa 8041 ctcgggcata aactctatga tttctctgga cgtggtgttg tcgaaactct caaagtacgc 8101 agtcaggaac gtgcgcgaca tgtcgtcggg aaactcgcgc ggaaacatgt tgttgtaacc 8161 gaacgggtcc catagcgcca aaaccaaatc tgccagcgtc aatagaatga gcacgatgcc 8221 gacaatggag ctggcttgga tagcgattcg agttaacgaa ttcttgaaga cgaaagggcc 8281 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct ta //