LOCUS Exported 5848 bp ds-DNA circular SYN 03-APR-2018 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5848) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 3, 2018 from SnapGene 4.1.7 http://www.snapgene.com COMMENT pHW539 15xUAS::kin-2a(G310D)::SL2::gfp::let-858 3'UTR PKA dominant negative effector construct the third GFP intron is corrected after sequencing with oHW233r G1.1 clone, the kin-2a(G310D)::SL2 section is confirmed by sequencing with oHW12f and oHW231r, largen 137213, at 20170515 Vector: pJL046 (15xUAS::HisCl-SL2::GFP::let-858 3'UTR) digested with KpnI and treated with CIP. ~ 4.7kb Insert: kin-2a(G310D, PKA-DN) PCR from pHW154(5.1) (for PKA dominant negative) with oHW435f+ oHW436r, ~1.1kb Gibson ligation. FEATURES Location/Qualifiers source 1..5848 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 36..52 /label=UAS /label=UAS(2) /note="color: #ff80c0" misc_feature 55..71 /label=UAS /label=UAS(8) /note="color: #ff80c0" misc_feature 74..90 /label=UAS /label=UAS(6) /note="color: #ff80c0" misc_feature 93..109 /label=UAS /label=UAS(12) /note="color: #ff80c0" misc_feature 112..117 /label=UAS /note="color: #ff80ff" misc_feature 119..128 /label=UAS /label=UAS(4) /note="color: #ff80ff" misc_feature 169..185 /label=UAS /label=UAS(15) /note="color: #ff80c0" misc_feature 188..204 /label=UAS /label=UAS(5) /note="color: #ff80c0" misc_feature 207..223 /label=UAS /label=UAS(7) /note="color: #ff80c0" misc_feature 226..242 /label=UAS /label=UAS(14) /note="color: #ff80c0" misc_feature 245..250 /label=UAS /label=UAS(10) /note="color: #ff80ff" misc_feature 252..261 /label=UAS /label=UAS(1) /note="color: #ff80ff" misc_feature 302..318 /label=UAS /label=UAS(17) /note="color: #ff80c0" misc_feature 321..337 /label=UAS /label=UAS(9) /note="color: #ff80c0" misc_feature 340..356 /label=UAS /label=UAS(13) /note="color: #ff80c0" misc_feature 359..375 /label=UAS /label=UAS(11) /note="color: #ff80c0" misc_feature 378..383 /label=UAS /label=UAS(16) /note="color: #ff80ff" misc_feature 385..394 /label=UAS /label=UAS(3) /note="color: #ff80ff" misc_feature 835..1935 /label=kin-2a(G310D) /note="color: #00ffff" misc_feature 1762..1764 /label=G310D mutation to abolish the binding of cAMP o /label=G310D mutation to abolish the binding of cAMP of Si /note="color: #ff0000" misc_feature 1945..2194 /label=SL2 /note="color: #ffff00" misc_feature 2204..2791 /label=GFP /note="color: #00ff00" misc_feature 2373..2422 /label=intron1 /note="color: #8080ff" misc_feature 2577..2627 /label=intron2 /note="color: #8080ff" misc_feature 2792..2842 /label=GFP 3intron corrected /note="color: #0000ff" misc_feature 2843..3070 /label=GFP /label=GFP(1) /note="color: #00ff00" misc_feature 3101..3533 /label=let-858 3UTR /label=nonstandard type: UTR /note="color: #ffc0cb" CDS 3896..4555 /label=AmpR /note="color: #ffff00; direction: RIGHT" rep_origin 4653..5335 /label=ColE1 origin /note="color: #808080" misc_binding 5675..5697 /label=LacO /note="color: #6495ed" primer_bind 5703..5723 /label=M13-rev /note="color: #00ffff; direction: RIGHT" intron 5738..5787 /label=decoy intron /note="color: #ff0acb" promoter 5788..2 /label=decoy ORF /note="color: #ff11ed" ORIGIN 1 aagcttgcta gcccatgatt acgccaagct acgggcggag tactgtcctc cgagcggagt 61 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact 121 gtcctccgag cggagactcc ctagcccatg attacgccaa gctacgggcg gagtactgtc 181 ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc 241 cgagcggagt actgtcctcc gagcggagac tccctagccc atgattacgc caagctacgg 301 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg 361 gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gactccctag gggcggccgc 421 actgactggg ccggccaaaa atgcataagg ttttgctggc actgttcttt atctttctgg 481 caccagcatc cgcactggca gtctccgaac cggcctgcag gtcgattttt tgcaaattac 541 gagcgttgta gggggcggag cgataggtcc tataggtttt ggtatatcat cattcattca 601 ttcattggta cattcattta cccaccttcc tctttctgag cttctctgga gttctgtgct 661 tcctttttcc cttatcttta tactgtaatt tttaactttc aggcattgat tggcgcgcct 721 ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat 781 aacatagaac attttcagga ggacccttgg ctagcgtcga cggtaccggt aaaaatgtcg 841 ggtggaaacg aagaggatca gttggcacaa tgccaggcct atgtgcaacg acacaacatt 901 cagcagcttg tcaaagaagc aattgttgtt ctgtgcatac acaaacccga caacccggtt 961 ctcttcttga aggatcattt tgaaaaactc aacgaacaac gagcacagga aggaggaaat 1021 ccggacgctg ctgatgatga cgacatcatt gtggagccac caaagagatc aggtggacgc 1081 agaaccggaa tctctgccga accaatcaag gaggacgata ccgagtacaa gaaggttgtg 1141 attccaaagg atgatgctac aagaaggtcg cttgagtcag ccatgaggaa gaacttgctc 1201 tttgctcatt tggaggagga tgagcaaaaa accatgtacg atgccatgtt cccagttgag 1261 aagtctgctg gagagaccat catcgaacag ggagaagaag gagataactt ctatgttatt 1321 gataagggaa ccgttgatgt ctacgtcaat cacgagtacg ttctcaccat caacgagggt 1381 ggatcatttg gagaacttgc tttgatctat ggaactccac gtgctgcaac cgtcattgcc 1441 aagaccgacg tgaaattgtg ggccatcgac cgtctcacct accgtcgtat cttgatggga 1501 tcagtaacaa agaagagaaa gatgtatgac gagttcctct ccaaggttca aatcttggcc 1561 gatctcgacc agtgggagcg cgctaacgtg gctgatgctc ttgagagatg cgacttcgag 1621 ccaggaactc atgtcgtcga gcaaggacaa cccggagacg aattcttcat cattctcgag 1681 ggagaagcca atgtgttgca gaagagaagc gacgacgcac cattcgacgt tgtcggacat 1741 cttggaatgt cggactattt cgacgaaatc gctcttcttc tcgaccgccc acgtgccgcc 1801 accgttgttg ccaaaaccca tctcaagtgc atcaagctgg atcgtaaccg cttcgaacgt 1861 gtcatgggac cagttcgtga gatcctgaag agagacgttt caaactacaa ctcatacgtc 1921 aaactgatga cctaaggtac cgtagctgtc tcatcctact ttcacctagt taactgcttg 1981 tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta caagtatata 2041 aatggtctct tctcaataaa ggttgtatat ttattcatct tattgaatct gccatttcct 2101 cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca acttctaatt 2161 ttaattcagg gaaactgctg taccggtaga aaaaatgctc gagagtaaag gagaagaact 2221 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt 2281 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat 2341 ttgcactact ggaaaactac ctgttccatg ggtaagttta aacatatata tactaactaa 2401 ccctgattat ttaaattttc agccaacact tgtcactact ttctgttatg gtgttcaatg 2461 cttctcgaga tacccagatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga 2521 aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtaa 2581 gtttaaacag ttcggtacta actaaccata catatttaaa ttttcaggtg ctgaagtcaa 2641 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga 2701 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat 2761 ggcagacaaa caaaagaatg gaatcaaagt tgtaagttta aacatgattt tactaactaa 2821 ctaatctgat ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt 2881 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc 2941 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga 3001 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact 3061 atacaaatag gaattcgcta gccggccata caagtaatcc ggatgatcga cgccaacgtc 3121 gttgaatttt caaattttaa atactgaata tttgtttttt ttcctattat ttatttattc 3181 tctttgtgtt ttttttcttg ctttctaaaa aattaattca atccaaatct aaacattttt 3241 ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag 3301 tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa 3361 acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc 3421 agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact 3481 agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct tgggcccaaa 3541 aggccttttt tttttggcgc cgacgtcagg tggcactttt cggggaaatg tgcgcggaac 3601 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 3661 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 3721 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 3781 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 3841 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 3901 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 3961 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 4021 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 4081 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 4141 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 4201 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 4261 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 4321 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 4381 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 4441 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 4501 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 4561 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 4621 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 4681 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 4741 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 4801 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 4861 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 5101 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga 5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 5341 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 5401 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 5461 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 5521 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 5581 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 5641 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 5701 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta 5761 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat 5821 ggatacgcta acaacttgga aatgaaat //