LOCUS Exported 5005 bp ds-DNA circular SYN 17-JUL-2006 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR221-P5P4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5005) AUTHORS Invitrogen TITLE Direct Submission JOURNAL Exported Jan 17, 2019 from SnapGene Viewer 4.2.11 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5005 /organism="synthetic DNA construct" /note="/dnas_title=osTIR1_P2A_mAID(fusion)" misc_feature complement(268..295) /label=rrnB T2 transcription termination sequence /note="rrnB T2 transcription termination sequence" misc_feature complement(427..470) /label=rrnB T1 transcription termination sequence /note="rrnB T1 transcription termination sequence" primer_bind 517..533 /label=M13 Forward (-40) primer binding site /note="M13 Forward (-40) primer binding site" primer_bind 537..552 /label=M13 Forward (-20) primer binding site /note="M13 Forward (-20) primer binding site" misc_recomb 570..665 /label=attL5 /note="attL5 recombination site" /note="recognized as attL5" gene 674..2827 /label=osTIR1 with NES and myc epitope tag(9x) /note="osTIR1 with NES and myc epitope tag(9x)" /note="derived from Addgene #47328" misc_feature 2471..2500 /label=myc epitope tag /note="myc epitope tag" misc_feature 2507..2536 /label=myc epitope tag /note="myc epitope tag" misc_feature 2543..2572 /label=myc epitope tag /note="myc epitope tag" misc_feature 2591..2620 /label=myc epitope tag /note="myc epitope tag" misc_feature 2627..2656 /label=myc epitope tag /note="myc epitope tag" misc_feature 2663..2692 /label=myc epitope tag /note="myc epitope tag" misc_feature 2711..2740 /label=myc epitope tag /note="myc epitope tag" misc_feature 2747..2776 /label=myc epitope tag /note="myc epitope tag" misc_feature 2783..2812 /label=myc epitope tag /note="myc epitope tag" misc_feature 2837..2845 /label=GSG linker /note="GSG linker" misc_feature 2846..2902 /label=P2A self-cleaving peptide /note="P2A self-cleaving peptide" misc_feature 2897..2902 /label=G^P cleavage site /note="G^P cleavage site" gene 2903..3106 /label=mAID /note="mAID" /note="Described in PMID: 27052166" misc_recomb complement(3117..3215) /label=attL4 /note="attL4 recombination site" /note="recognized as attL4" misc_feature join(3127..5005,1..658) /label=nonstandard type: vector /note="/source=pDONR221-P5P4" /note="/type=Donor Vector" primer_bind complement(3270..3286) /label=M13 Reverse primer binding site /note="M13 Reverse primer binding site" gene 3399..4208 /label=Kanamycin resistance gene /note="Kanamycin resistance gene" rep_origin 4329..5002 /label=f1 origin /note="f1 origin" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa 661 agttgtcgcc accatggcat tacaaaaaaa attagaagaa ttagaattag atgaaggcgg 721 cacgtacttc ccggaggagg tggtggagca catcttcagc ttcctgccgg cgcagcgcga 781 ccgcaacacg gtctcgctcg tctgcaaggt gtggtacgag atcgagaggc tgagccgccg 841 cggcgtcttc gtgggcaact gctacgccgt gcgcgccggc cgcgtcgccg cgcggttccc 901 caacgtgcgg gcgctcacgg tgaaggggaa gccccacttc gccgacttca acctcgtgcc 961 ccccgactgg ggcggctacg cggggccgtg gatcgaggcg gccgcgaggg gatgccacgg 1021 cctggaggag ctcaggatga agcggatggt ggtgtccgac gagagcctcg agctgctggc 1081 tcgctcgttc ccgcggttca gggctcttgt tcttatcagc tgcgaggggt tcagcactga 1141 cgggctagcc gccgtcgcga gccattgcaa gcttctgagg gagttggatt tgcaggaaaa 1201 tgaagtggag gatcgagggc ctaggtggct ttcctgcttc cctgattcct gcacatcact 1261 tgtctcattg aattttgcct gcatcaaagg ggaggttaat gctggttcac tggagagact 1321 tgttagcagg tccccaaacc tgcggagttt gaggctgaat cgatctgtat cggtagatac 1381 acttgcaaag atactactgc gtacccctaa cttggaggat ttggggacag ggaatttgac 1441 agatgacttc caaactgagt cctactttaa gcttaccagt gctctggaga aatgcaagat 1501 gttgaggagt ttgtctggat tctgggatgc ttctcctgtt tgcctgtcat ttatctaccc 1561 cctgtgtgct caactgacag gattgaactt gagctatgca cccacacttg atgcttctga 1621 ccttacaaaa atgattagcc gctgtgtgaa gctccaacgc ctttgggtac tggattgtat 1681 ctcggacaaa ggcttgcaag tggtggcctc cagttgcaaa gacttgcaag aactcagggt 1741 atttccatca gatttctacg tagctggtta ttctgcagtg acagaggagg gacttgttgc 1801 agtatccttg ggctgtccaa aactgaactc actactgtac ttctgtcacc aaatgactaa 1861 tgctgcacta gttactgtcg ccaagaactg tccaaatttc acacgattca gactttgtat 1921 tcttgagcca gggaagcctg atgttgtgac aagccaacca ttagatgaag gctttggagc 1981 tattgttcgt gagtgcaagg gattacaacg tttgtcaata tctggtcttc tcacagacaa 2041 agttttcatg tatatcggga aatatgcaaa acaacttgag atgctttcta tagcatttgc 2101 tggtgacagt gataagggta tgatgcatgt tatgaatgga tgcaagaatt taaggaaact 2161 ggagataaga gatagcccgt ttggtgatgc tgcactcttg gggaattttg ctaggtacga 2221 gacaatgcga tccctttgga tgtcatcttg caatgtcacg ttaaaggggt gccaagtcct 2281 tgcgtcaaag atgccgatgc tcaatgttga ggtcataaat gagcgggatg gtagcaatga 2341 aatggaggaa aaccatggag atctgcctaa agtggagaaa ttatatgtgt accgcacaac 2401 tgctggggcg agggatgatg caccaaattt tgttaaaatc ctagtcgact ccggttctgc 2461 tgctagtggt gaacaaaagt tgatttctga agaagatttg aacggtgaac aaaagctaat 2521 ctccgaggaa gacttgaacg gtgaacaaaa attaatctca gaagaagact tgaacggatc 2581 ctctagaggt gaacaaaagt tgatttctga agaagatttg aacggtgaac aaaagctaat 2641 ctccgaggaa gacttgaacg gtgaacaaaa attaatctca gaagaagact tgaacggatc 2701 ctctagaggt gaacaaaagt tgatttctga agaagatttg aacggtgaac aaaagctaat 2761 ctccgaggaa gacttgaacg gtgaacaaaa attaatctca gaagaagact tgaacggatc 2821 cactagcggt acagccggat caggagctac taatttctct ctgttaaagc aggcaggaga 2881 tgtggaggag aatccaggtc ctaaggagaa gagtgcttgt cctaaagatc cagccaaacc 2941 tccggccaag gcacaagttg tgggatggcc accggtgaga tcataccgga agaacgtgat 3001 ggtttcctgc caaaaatcaa gcggtggccc ggaggcggcg gcgttcgtga aggtatcaat 3061 ggacggagca ccgtacttga ggaaaatcga tttgaggatg tataaaggca gtaccaaccc 3121 aacttttcta tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga 3181 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ggcggccgcg 3241 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg 3301 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa 3361 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa 3421 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc 3481 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc 3541 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg 3601 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta 3661 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat 3721 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc 3781 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg 3841 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc 3901 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac 3961 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga 4021 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg 4081 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa 4141 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt 4201 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac 4261 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag 4321 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 4381 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 4441 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 4501 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 4561 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 4621 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 4681 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 4741 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 4801 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 4861 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 4921 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 4981 tttgctggcc ttttgctcac atgtt //