LOCUS Exported 8493 bp ds-DNA circular SYN 17-JUL-2006 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR221-P4rP3r SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8493) 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..8493 /organism="synthetic DNA construct" /note="/dnas_title=IF301_dSaCas9_LSD1" 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 complement(570..726) /label=attL4r /note="attL4r recombination site" /note="recognized as attR4" misc_feature 732..782 /label=NLS /note="NLS" gene 783..3938 /label=hSaCas9 (D10A, N580A) /note="hSaCas9 (D10A, N580A)" /note="nuclease deficient" misc_feature 1463..1481 /label=nonstandard type: PCR_primer /note="JH256 sequencing primer" misc_feature 2339..2355 /label=nonstandard type: PCR_primer /note="JH257 sequencing primer" misc_feature 3177..3194 /label=nonstandard type: PCR_primer /note="JH258 sequencing primer" misc_feature 3866..3884 /label=nonstandard type: PCR_primer /note="JH259 sequencing primer" misc_feature 3939..3986 /label=NLS /note="NLS" misc_feature 3987..3998 /label=GGGS linker /note="GGGS linker" gene 3999..6551 /label=LSD1 /note="LSD1" misc_feature 4593..4610 /label=nonstandard type: PCR_primer /note="JH325 seq primer" misc_feature 5223..5240 /label=nonstandard type: PCR_primer /note="JH326 seq primer" misc_recomb 6556..6712 /label=attL3r /note="attL3r recombination site" /note="recognized as attR3" misc_feature join(6563..8493,1..719) /label=nonstandard type: vector /note="/source=pDONR221-P4rP3r" /note="/type=Donor Vector" primer_bind complement(6758..6774) /label=M13 Reverse primer binding site /note="M13 Reverse primer binding site" gene 6887..7696 /label=Kanamycin resistance gene /note="Kanamycin resistance gene" rep_origin 7817..8490 /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 tcgggcccct acaggtcact aataccatct aagtagttga 601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa 661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca 721 aagttgccac catggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag 781 ccaagcggaa ctacatcctg ggcctggcca tcggcatcac cagcgtgggc tacggcatca 841 tcgactacga gacacgggac gtgatcgatg ccggcgtgcg gctgttcaaa gaggccaacg 901 tggaaaacaa cgagggcagg cggagcaaga gaggcgccag aaggctgaag cggcggaggc 961 ggcatagaat ccagagagtg aagaagctgc tgttcgacta caacctgctg accgaccaca 1021 gcgagctgag cggcatcaac ccctacgagg ccagagtgaa gggcctgagc cagaagctga 1081 gcgaggaaga gttctctgcc gccctgctgc acctggccaa gagaagaggc gtgcacaacg 1141 tgaacgaggt ggaagaggac accggcaacg agctgtccac caaagagcag atcagccgga 1201 acagcaaggc cctggaagag aaatacgtgg ccgaactgca gctggaacgg ctgaagaaag 1261 acggcgaagt gcggggcagc atcaacagat tcaagaccag cgactacgtg aaagaagcca 1321 aacagctgct gaaggtgcag aaggcctacc accagctgga ccagagcttc atcgacacct 1381 acatcgacct gctggaaacc cggcggacct actatgaggg acctggcgag ggcagcccct 1441 tcggctggaa ggacatcaaa gaatggtacg agatgctgat gggccactgc acctacttcc 1501 ccgaggaact gcggagcgtg aagtacgcct acaacgccga cctgtacaac gccctgaacg 1561 acctgaacaa tctcgtgatc accagggacg agaacgagaa gctggaatat tacgagaagt 1621 tccagatcat cgagaacgtg ttcaagcaga agaagaagcc caccctgaag cagatcgcca 1681 aagaaatcct cgtgaacgaa gaggatatta agggctacag agtgaccagc accggcaagc 1741 ccgagttcac caacctgaag gtgtaccacg acatcaagga cattaccgcc cggaaagaga 1801 ttattgagaa cgccgagctg ctggatcaga ttgccaagat cctgaccatc taccagagca 1861 gcgaggacat ccaggaagaa ctgaccaatc tgaactccga gctgacccag gaagagatcg 1921 agcagatctc taatctgaag ggctataccg gcacccacaa cctgagcctg aaggccatca 1981 acctgatcct ggacgagctg tggcacacca acgacaacca gatcgctatc ttcaaccggc 2041 tgaagctggt gcccaagaag gtggacctgt cccagcagaa agagatcccc accaccctgg 2101 tggacgactt catcctgagc cccgtcgtga agagaagctt catccagagc atcaaagtga 2161 tcaacgccat catcaagaag tacggcctgc ccaacgacat cattatcgag ctggcccgcg 2221 agaagaactc caaggacgcc cagaaaatga tcaacgagat gcagaagcgg aaccggcaga 2281 ccaacgagcg gatcgaggaa atcatccgga ccaccggcaa agagaacgcc aagtacctga 2341 tcgagaagat caagctgcac gacatgcagg aaggcaagtg cctgtacagc ctggaagcca 2401 tccctctgga agatctgctg aacaacccct tcaactatga ggtggaccac atcatcccca 2461 gaagcgtgtc cttcgacaac agcttcaaca acaaggtgct cgtgaagcag gaagaagcca 2521 gcaagaaggg caaccggacc ccattccagt acctgagcag cagcgacagc aagatcagct 2581 acgaaacctt caagaagcac atcctgaatc tggccaaggg caagggcaga atcagcaaga 2641 ccaagaaaga gtatctgctg gaagaacggg acatcaacag gttctccgtg cagaaagact 2701 tcatcaaccg gaacctggtg gataccagat acgccaccag aggcctgatg aacctgctgc 2761 ggagctactt cagagtgaac aacctggacg tgaaagtgaa gtccatcaat ggcggcttca 2821 ccagctttct gcggcggaag tggaagttta agaaagagcg gaacaagggg tacaagcacc 2881 acgccgagga cgccctgatc attgccaacg ccgatttcat cttcaaagag tggaagaaac 2941 tggacaaggc caaaaaagtg atggaaaacc agatgttcga ggaaaagcag gccgagagca 3001 tgcccgagat cgaaaccgag caggagtaca aagagatctt catcaccccc caccagatca 3061 agcacattaa ggacttcaag gactacaagt acagccaccg ggtggacaag aagcctaata 3121 gagagctgat taacgacacc ctgtactcca cccggaagga cgacaagggc aacaccctga 3181 tcgtgaacaa tctgaacggc ctgtacgaca aggacaatga caagctgaaa aagctgatca 3241 acaagagccc cgaaaagctg ctgatgtacc accacgaccc ccagacctac cagaaactga 3301 agctgattat ggaacagtac ggcgacgaga agaatcccct gtacaagtac tacgaggaaa 3361 ccgggaacta cctgaccaag tactccaaaa aggacaacgg ccccgtgatc aagaagatta 3421 agtattacgg caacaaactg aacgcccatc tggacatcac cgacgactac cccaacagca 3481 gaaacaaggt cgtgaagctg tccctgaagc cctacagatt cgacgtgtac ctggacaatg 3541 gcgtgtacaa gttcgtgacc gtgaagaatc tggatgtgat caaaaaagaa aactactacg 3601 aagtgaatag caagtgctat gaggaagcta agaagctgaa gaagatcagc aaccaggccg 3661 agtttatcgc ctccttctac aacaacgatc tgatcaagat caacggcgag ctgtatagag 3721 tgatcggcgt gaacaacgac ctgctgaacc ggatcgaagt gaacatgatc gacatcacct 3781 accgcgagta cctggaaaac atgaacgaca agaggccccc caggatcatt aagacaatcg 3841 cctccaagac ccagagcatt aagaagtaca gcacagacat tctgggcaac ctgtatgaag 3901 tgaaatctaa gaagcaccct cagatcatca aaaagggcaa aaggccggcg gccacgaaaa 3961 aggccggcca ggcaaaaaag aaaaagggag gaggatcttt atctgggaag aaggcggcag 4021 ccgcggcggc ggcggctgca gcggcagcaa ccgggacgga ggctggccct gggacagcag 4081 gcggctccga gaacgggtct gaggtggccg cgcagcccgc gggcctgtcg ggcccagccg 4141 aggtcgggcc gggggcggtg ggggagcgca caccccgcaa gaaagagcct ccgcgggcct 4201 cgccccccgg gggcctggcg gaaccgccgg ggtccgcagg gcctcaggcc ggccctactg 4261 tcgtgcctgg gtctgcgacc cccatggaaa ctggaatagc agagactccg gaggggcgtc 4321 ggaccagccg gcgcaagcgg gcgaaggtag agtacagaga gatggatgaa agcttggcca 4381 acctctcaga agatgagtat tattcagaag aagagagaaa tgccaaagca gagaaggaaa 4441 agaagcttcc cccaccaccc cctcaagccc cacctgagga agaaaatgaa agtgagcctg 4501 aagaaccatc gggtgtggag ggcgcagctt tccagagccg acttcctcat gaccggatga 4561 cttctcaaga agcagcctgt tttccagata ttatcagtgg accacaacag acccagaagg 4621 tttttctttt cattagaaac cgcacactgc agttgtggtt ggataatcca aagattcagc 4681 tgacatttga ggctactctc caacaattag aagcacctta taacagtgat actgtgcttg 4741 tccaccgagt tcacagttat ttagagcgtc atggtcttat caacttcggc atctataaga 4801 ggataaaacc cctaccaact aaaaagacag gaaaggtaat tattataggc tctggggtct 4861 caggcttggc agcagctcga cagttacaaa gttttggaat ggatgtcaca cttttggaag 4921 ccagggatcg tgtgggtgga cgagttgcca catttcgcaa aggaaactat gtagctgatc 4981 ttggagccat ggtggtaaca ggtcttggag ggaatcctat ggctgtggtc agcaaacaag 5041 taaatatgga actggccaag atcaagcaaa aatgcccact ttatgaagcc aacggacaag 5101 ctgttcctaa agagaaagat gaaatggtag agcaagagtt taaccggttg ctagaagcta 5161 catcttacct tagtcatcaa ctagacttca atgtcctcaa taataagcat gtgtcccttg 5221 gccaggcatt ggaagttgtc attcagttac aagagaagca tgtcaaagat gagcagattg 5281 aacattggaa gaagatagtg aaaactcagg aagaattgaa agaacttctt aataagatgg 5341 taaatttgaa agagaaaatt aaagaactcc atcagcaata caaagaagca tctgaagtaa 5401 agccacccag agatattact gccgagttct tagtgaaaag caaacacagg gatctgaccg 5461 ccctatgcaa ggaatatgat gaattagctg aaacacaagg aaagctagaa gaaaaacttc 5521 aggagttgga agcgaatccc ccaagtgatg tatatctctc atcaagagac agacaaatac 5581 ttgattggca ttttgcaaat cttgaatttg ctaatgccac acctctctca actctctccc 5641 ttaagcactg ggatcaggat gatgactttg agttcactgg cagccacctg acagtaagga 5701 atggctactc gtgtgtgcct gtggctttag cagaaggcct agacattaaa ctgaatacag 5761 cagtgcgaca ggttcgctac acggcttcag gatgtgaagt gatagctgtg aatacccgct 5821 ccacgagtca aacctttatt tataaatgcg acgcagttct ctgtaccctt cccctgggtg 5881 tgctgaagca gcagccacca gccgttcagt ttgtgccacc tctccctgag tggaaaacat 5941 ctgcagtcca aaggatggga tttggcaacc ttaacaaggt ggtgttgtgt tttgatcggg 6001 tgttctggga tccaagtgtc aatttgttcg ggcatgttgg cagtacgact gccagcaggg 6061 gtgagctctt cctcttctgg aacctctata aagctccaat actgttggca ctagtggcag 6121 gagaagctgc tggtatcatg gaaaacataa gtgacgatgt gattgttggc cgatgcctgg 6181 ccattctcaa agggattttt ggtagcagtg cagtacctca gcccaaagaa actgtggtgt 6241 ctcgttggcg tgctgatccc tgggctcggg gctcttattc ctatgttgct gcaggatcat 6301 ctggaaatga ctatgattta atggctcagc caatcactcc tggcccctcg attccaggtg 6361 ccccacagcc gattccacga ctcttctttg cgggagaaca tacgatccgt aactacccag 6421 ccacagtgca tggtgctctg ctgagtgggc tgcgagaagc gggaagaatt gcagaccagt 6481 ttttgggggc catgtatacg ctgcctcgcc aggccacacc aggtgttcct gcacagcagt 6541 ccccaagcat gggcgcaact ttgtataata aagttgaacg agaaacgtaa aatgatataa 6601 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac 6661 aacatatcca gtcactatga atcaactact tagatggtat tagtgacctg tactgcaggg 6721 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct 6781 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg 6841 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca 6901 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa 6961 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc 7021 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga 7081 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt 7141 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt 7201 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt 7261 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt 7321 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga 7381 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc 7441 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga 7501 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca 7561 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct 7621 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct 7681 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct 7741 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt 7801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 7861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 7921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 7981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 8041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 8101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 8161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 8221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 8281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 8341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 8401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 8461 cctggccttt tgctggcctt ttgctcacat gtt //