LOCUS Exported 10005 bp ds-DNA circular SYN 06-MAY-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10005) AUTHORS . TITLE Direct Submission JOURNAL Exported May 6, 2018 from SnapGene Viewer 4.0.7 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10005 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 860..951 /label=Amp Promoter /note="Amp Promoter" misc_structure 1232..1254 /label=AmpR CrispR target /note="AmpR CrispR target" misc_structure 1255..2294 /label=Homology Arm Left (HAL4) /note="Homology Arm Left (HAL4)" misc_structure 2295..2302 /label=MCS Site 1 (AarI flanked) [Split] /note="MCS Site 1 (AarI flanked) [Split]" misc_structure 2303..2305 /label=MCS Site 1 (AarI flanked) [Split] /note="MCS Site 1 (AarI flanked) [Split]" misc_RNA 2306..2333 /note="Build in of NdeI site for cloning into pHD-dAttP-3xP3DsRed-HAR1_HAL4_AmpR(AttP40)" misc_feature 2334..3092 /note="20X UAS + HSP70 minimal promoter from pJFRC7-20XUAS-IVS-mCD8-GFP" misc_feature 3140..3146 /label=Drosophila Kozak Sequence /note="Drosophila Kozak Sequence" gene 3161..3194 /label=AttP34 according to Groth et al /note="AttP34 according to Groth et al" intron 3298..3304 /note="small T Intron from pJFRC210-10XUAS-FRT>STOP>FRT-myr::smGFP-OLLAS Note - This was taken from Nern et al., 2015 (PNAS) [Split]" intron 3305..3363 /note="small T Intron from pJFRC210-10XUAS-FRT>STOP>FRT-myr::smGFP-OLLAS Note - This was taken from Nern et al., 2015 (PNAS) [Split]" misc_feature 3493..3513 /note="SV40 NLS from pJFRC210-10XUAS-FRT>STOP>FRT-myr::smGFP-OLLAS" misc_feature 3514..3762 /label=In line with SV40 NLS /note="In line with SV40 NLS" misc_feature 3785..3919 /label=SV40 Poly A /note="SV40 Poly A" gene 3931..3990 /label=AttP Wt according to Groth et al /note="AttP Wt according to Groth et al" gene 4322..4391 /label=AttBWT according to Groth et al., 2000 /note="AttBWT according to Groth et al., 2000" misc_feature 4398..5822 /label=LexA-p65 fusion? /note="LexA-p65 fusion?" misc_feature 4398..5003 /label=LexA DNA binding domain /note="LexA DNA binding domain" misc_feature 4398..5003 /label=LexA DNA binding domain /note="LexA DNA binding domain" misc_feature 5004..5015 /label=Linker /note="Linker" misc_feature 5016..5819 /label=p65 activation domain /note="p65 activation domain" misc_feature 5837..6097 /label=hsp70 3'UTR /note="hsp70 3'UTR" misc_feature 5837..6097 /label=Hsp70 3' Terminator /note="Hsp70 3' Terminator" gene 5837 /label=LexA [Split] /note="LexA [Split]" misc_RNA 6098..6162 /note="Build back of LoxP site for BsiWI cloning into pHD-dAttP-3xP3DsRed-HAR1_HAL4_AmpR(AttP40)" misc_feature 6163..6368 /label=3xP3 Enhancer + hsp70 min /note="3xP3 Enhancer + hsp70 min" gene 6388..7065 /label=DsRed /note="DsRed" misc_recomb 7314..7347 /label=LoxP Site 2 /note="LoxP Site 2" misc_structure 7348..8538 /label=HAR1 for AttP40 /note="HAR1 for AttP40" misc_structure 8516..8538 /label=AmpR gRNA CRISPR Target /note="AmpR gRNA CRISPR Target" oriT 9292..9880 ORIGIN 1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc 181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt 901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca 961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg 1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg 1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga 1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg 1201 cccgggctaa ttatggggtg tcgcccttcg cacagaaaag catcttacgg atggtgtttg 1261 ccaaaagtgc tggagatgtt ttagatatag atggtttatt ttaagaaatg gatgtatata 1321 attgagctac tgcatcacgg gtttctgttt aacagtaaca gtgaaaagtg ctcaatgata 1381 catatattac aaataataac ttttggtttt taaagtagat ttatataata taatcataat 1441 tcttaatggg aatccatgtt acgaccaaat tagccgtggc ctttcaagtg gctgccactt 1501 atattttaca actcgcacac agtggctcac accctcgtaa acaaaataat gctattaaat 1561 caaattgaac atgtgaattc tctcaagaag acgccacttt tttttggggg gccagctgca 1621 actgccacca atgtttactt ttatctttaa tgggcacatg ccttacatat acgaatttta 1681 cacgattttt caagtggcgg tcgagagtgg ggtgataatt aattaaggtt gcctttagat 1741 aaagtattat agagtttcaa aaaaatgaaa cttaagaatg gaatagctgc attccattct 1801 atgtagattt cccttcaaaa ttattaaaat ccgttgacct ttgttcattt agcaattaga 1861 attcgtctat gaataacaaa ttaaattgcg ttgcgggcaa attgctttag gccatttgaa 1921 gtgcaacttt atatctcagg gccaacttct cggcgcgaca acaataaaaa catatacata 1981 gatacacata tcagatgcaa tgcaatcacc acagcgtgac gtcattagtc taggcttgaa 2041 aatacacggc aaacttgttt ataactctac gctgattaca taagccgcaa acacgacgaa 2101 aaaacatttg ttttggtttc aaatccatga atgactttca ttaagtgcag ctgtaagctg 2161 agctttgcca tgcgctgacc catattttca gcctctcgtt ctctctgttt cgcagatact 2221 gtgaatatta gtaaaacgat ccataggggc tgtagtttat aatcataatc aaataggcat 2281 aaacaaccac cagcgcggcc gcggacatat gggatccagt gcaccgtaag cttgcatgcc 2341 tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc 2401 cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagact ctagccctag 2461 ggcatgcctg caggtcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta 2521 ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagactct 2581 agcgctagcg catgcctgca ggtcggagta ctgtcctccg agcggagtac tgtcctccga 2641 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg 2701 gagactctag cactagtgca tgcctgcagg tcggagtact gtcctccgag cggagtactg 2761 tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc 2821 tccgagcgga gactctagcg acgtcgagcg ccggagtata aatagaggcg cttcgtctac 2881 ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa 2941 ataaacaagc gcagctgaac aagctaaaca atctgcagta aagtgcaagt taaagtgaat 3001 caattaaaag taaccagcaa ccaagtaaat caactgcaac tactgaaatc tgccaagaag 3061 taattattga atacaagaag agaactctga attaacgtaa gctagctaga ggatctttgt 3121 gaaggaacct tacggtacca atcaaaagat ctactccagt ccaactgggg taacctttga 3181 gttctctcag ttggtaatac gtaagctagc tagaggatct ttgtgaagga accttacttg 3241 gtaccctgtg gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta 3301 aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt 3361 tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa 3421 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa 3481 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa 3541 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt 3601 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atatttgatg 3661 tatagtgcct tgactagaga tcataatcag ccataccaca tttgtagagg ttttacttgc 3721 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 3781 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3901 atcttatcat gtctggatcg atctggccgg cgggagtagt gccccaactg gggtaacctt 3961 tgagttctct cagttggggg cgtagggtcg gggtcggcat ggcatctcca cctcctcgcg 4021 gtccgacctg ggcatccgaa ggaggacgtc gtccactcgg atggctaagg gagagctcgg 4081 atccgggcaa tacgaggtac tggcagacac gcagtgtcga acaacacctg acctaacggt 4141 aagatgagta gataactcaa ctacttaaat cctggcctgc ttgatatctc gtagcgttct 4201 tagagatcga cgaactgttt cacgaccggg tcggcatggc atctccacct cctcgcggtc 4261 cgacctgggc atccgaagga ggacgtcgtc cactcggatg gctaagggag agctcggatc 4321 cctcgaagcc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca 4381 cctcacccat cgtcgacatg aaggctctca cggcccgaca acaggaagtt tttgatttga 4441 tacgggatca tatatcccaa acgggtatgc ctccgacccg cgcagagata gcacagcgac 4501 tgggctttcg atcgcctaac gccgcggagg agcacttgaa ggcactggcc cgcaagggtg 4561 tcattgaaat cgtgtccggt gcgagccgcg gaatccggct gttgcaggag gaggaagagg 4621 gcctgccact ggtgggacgc gtggccgctg gcgagccgct gctggcccag caacacatag 4681 agggacacta tcaggtggac ccctccttgt ttaagccaaa tgctgatttc ctgttgcggg 4741 tgtcgggaat gtccatgaag gacatcggta ttatggatgg tgacctcctc gccgtccata 4801 agacacagga tgtcaggaac ggccaggtag tcgttgccag gatagacgat gaggtcactg 4861 tgaaacgtct caagaagcaa ggcaataagg tcgagctgct gccggagaat agcgagttca 4921 agccgatcgt ggtggatctg cgacagcagt cctttactat cgagggcttg gccgtgggtg 4981 tgatccgcaa cggagattgg ctgggatcca cgccgatgga gttccagtac ctgcccgata 5041 cggatgaccg tcaccgtatc gaagaaaagc ggaagcgaac ctatgaaacc ttcaagtcca 5101 tcatgaaaaa gtcccccttc tcgggcccca cggacccgcg ccccccgccc cgtcgtattg 5161 cggttccttc gcgcagcagt gccagcgtcc ccaaacccgc accgcagccc tacccgttca 5221 cttcctccct tagcacgatt aactatgatg agttccccac gatggtgttc cccagtggac 5281 aaatttccca ggcatcggca ctggctccgg ccccaccgca agtcctcccc caggcgcccg 5341 ctccggcacc ggctcccgca atggtgagtg ctctggccca ggcccccgct ccagtccccg 5401 tgctggcgcc tggaccccca caggcagttg cccctcctgc tccgaaacca acgcaggcgg 5461 gcgaaggaac cctgagcgag gccctcttgc agcttcagtt cgatgacgaa gacttgggag 5521 ccctgctggg taacagcaca gaccctgccg tattcaccga tctcgcatcc gtggacaaca 5581 gcgagtttca gcagctcttg aatcagggaa tcccggtcgc acctcatacc acagagccca 5641 tgctgatgga atacccggag gctatcacgc gactggtgac cggcgcacag cgaccacccg 5701 atccagcccc tgccccactg ggtgccccgg gtttgcccaa tggcctcctc agcggcgatg 5761 aggatttctc cagcatcgct gatatggatt tctccgcttt gctgagccag ataagctcct 5821 aagtttaaac aagcttatcg ataccgtcga ctaaagccaa atagaaaatt attcagttcc 5881 tggcttaagt ttttaaaagt gatattattt atttggttgt aaccaaccaa aagaatgtaa 5941 ataactaata cataattatg ttagttttaa gttagcaaca aattgatttt agctatatta 6001 gctacttggt taataaatag aatatattta tttaaagata attgcgtttt tattgtcagg 6061 gagtgagttt gcttaaaaac tcgtttagat ccactaggtc gactgaggca ctgcctcgag 6121 agataacttc gtataatgta tgctatacga agttatcgta cgggatctaa ttcaattaga 6181 gactaattca attagagcta attcaattag gatccaagct tatcgatttc gaaccctcga 6241 ccgccggagt ataaatagag gcgcttcgtc tacggagcga caattcaatt caaacaagca 6301 aagtgaacac gtcgctaagc gaaagctaag caaataaaca agcgcagctg aacaagctaa 6361 acaatcggct cgaagccggt cgccaccatg gcctcctccg aggacgtcat caaggagttc 6421 atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 6481 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc 6541 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg 6601 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc 6661 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 6721 tccctccagg acggctcctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc 6781 gacggccccg taatgcagaa gaagactatg ggctgggagg cgtccaccga gcgcctgtac 6841 ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa ggacggcggc 6901 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc 6961 tactactacg tggactccaa gctggacatc acctcccaca acgaggacta caccatcgtg 7021 gagcagtacg agcgcgccga gggccgccac cacctgttcc tgtaggggcc gcgactctag 7081 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 7141 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 7201 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 7261 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacc ggtataactt 7321 cgtataatgt atgctatacg aagttatcac attgggtagc acagactctg agctttacta 7381 cgattactat acagctcttc tctcgcgact ttttggactg gacaaggcgt agcacattga 7441 acggcagtgg gtttgggttt agtatcgaac cggctttcta cgacagcgga ttggaagcgc 7501 ggagcgacaa agtcgccggg taataatgtg atatagccgg ctatgttcag caggcaaaac 7561 tgaaataaaa gtaataaaca ccgaaagccc ccgaattctc gcatcttctt ttgtgggact 7621 gcgagaagaa ttgcgagcga aatactaatt tccttaccta actgtgtgag ataatctccg 7681 gttggcattg tgtgtcgcgc aataaaatcc ccaaaagccc aaagcttcaa gtgtttaata 7741 taataaaata gccaagaagc gtgtgtattt gcatttggga aaaatttaag gttgaattac 7801 gtgggccgaa aaattaaaga ctgaaggcca ttcaagggtt gttcaaagag aaaattcggt 7861 gaaaagtcac cgtgtgtgtg tgtgtgagtg tgtgtgcgtt gcgtgagaaa aatcaattct 7921 gcatttggtg gaaagccgaa aacggagaga aaattggtgg agcgaggaga aaaggacacg 7981 aattaaaggc tggttttttg gggctgcgaa aaactcgtga ccgatgacaa tttggcttga 8041 cgagggtcta ttagcgaaat caatgctatt atggctgata gtggaggacc aggatccaag 8101 cacatggatc caacaaccat agatgccggc gggggcggag caggagcagc cggaggtgat 8161 gacgtgcctc cagtcccagc tgttcgccgg cgtcgtgcgc atgagcagaa atcctcgaga 8221 gagcaggtgc tggaggagga gaagagccaa cagctggaga catcgacgat gacgaggacc 8281 tacatgaaga ccatcaccac ctccctcacc acctcgagta gctccaacgt ggaagagttt 8341 atcctggggg agcacgcagc aggagcagca gctgctccat cgcccaacca gcagcgattg 8401 cgtcaaaagg tggcccagta cgagaaggtc tggtcggatg gctccagtcc cgttaagcgt 8461 ccagccgagc agagctctga tgagctgcgt ctgaccgacg accaggatga gtacgccatc 8521 cgtaagatgc ttttctgtgt caatcgagtt caagggcgac acaaaattta ttctaaatgc 8581 ataataaata ctgataacat cttatagttt gtattatatt ttgtattatc gttgacatgt 8641 ataattttga tatcaaaaac tgattttccc tttattattt tcgagattta ttttcttaat 8701 tctctttaac aaactagaaa tattgtatat acaaaaaatc ataaataata gatgaatagt 8761 ttaattatag gtgttcatca atcgaaaaag caacgtatct tatttaaagt gcgttgcttt 8821 tttctcattt ataaggttaa ataattctca tatatcaagc aaagtgacag gcgcccttaa 8881 atattctgac aaatgctctt tccctaaact ccccccataa aaaaacccgc cgaagcgggt 8941 ttttacgtta tttgcggatt aacgattact cgttatcaga accgcccagg gggcccgagc 9001 ttaagactgg ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa aaaggccatc 9061 cgtcaggggc cttctgctta gtttgatgcc tggcagttcc ctactctcgc cttccgcttc 9121 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 9181 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 9241 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 9301 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 9361 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 9421 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 9481 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 9541 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 9601 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 9661 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggg ctaactacgg 9721 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 9781 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 9841 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 9901 tacggggtct gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg gattttggtc 9961 atgagcttgc gccgtcccgt caagtcagcg taatgctctg ctttt //