LOCUS Addgene62451 9171 bp DNA circular UNA 13-APR-2015 DEFINITION ES-FUCCI. ACCESSION urn.local...1420721390042.43 VERSION urn.local...1420721390042.43 KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers rep_origin 1153..1741 /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" sig_peptide 122..190 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein." /label="bla signal peptide" misc_RNA 1189..1741 /note="RNAII transcript (complementary strand)" /label="RNA" misc_RNA complement(1192..1299) /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(1308..1313) /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" -10_signal 73..78 /note="bla promoter (counter-clockwise); GAGACA" /label="-10 signal" -10_signal 1174..1179 /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(1330..1335) /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -35_signal 52..57 /note="bla promoter (counter-clockwise); TTCAAA" /label="-35 signal" -35_signal 1153..1158 /note="RNAII promoter (counter-clockwise); TTGAGA" /label="-35 signal" polyA_signal complement(<1921..3088) /label="beta-Globin poly A signal" polyA_signal complement(<4653..>4877) /label="bGHpA" polyA_signal <8941..>9165 /label="bGHpA" promoter complement(5897..6404) /label="PGK" promoter complement(4140..4647) /label="PGK" promoter 7659..7864 /label="minPGK" misc_feature complement(<7335..7636) /label="PGKenhancer" misc_feature complement(<7027..7328) /label="PGKenhancer" misc_feature complement(<6719..7020) /label="PGKenhancer" misc_feature complement(<6411..6712) /label="PGKenhancer" gene complement(3100..4122) /label="HygroR" gene 122..982 /gene="bla" /label="bla gene" gene 7879..8594 /label="Citrine" gene complement(5172..5882) /label="mCherry" gene 8602..8931 /label="hGeminin(1-110)" gene complement(4887..5159) /label="hCdt1(30-120)" ORIGIN 1 aattcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 61 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 1021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 1081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 1141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 1201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 1261 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 1321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 1381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 1441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 1501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 1561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 1621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 1681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 1741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 1801 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 1861 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcc atgagtcgac 1921 gcagtcgagt tcataagaga agagggacag ctatgactgg gagtagtcag gagaggagga 1981 aaaatctggc tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca 2041 caaaacaaaa tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat 2101 ggacagcagg gggctgtttc atatactgat gacctcttta tagccacctt tgttcatggc 2161 agccagcata tggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa 2221 tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattccc acacactatt 2281 gcaatgaaaa taaatttcct ttattaccca gaagtcagat gctcaagggg cttcatgatg 2341 tccccataat ttttggcaga gggaaaaaga tctcagtggt atttgtgagc cagggcattg 2401 gccacaccag ccaccacctt ctgataggca gcctgcacct gaggagtgaa ttctttgcca 2461 aaatgatgag acagcacaac aaccagcacg ttgcccagga gctgtaggaa aaagaagaag 2521 gcatgaacat ggttagcaga ggggcccggt ttggactcag agtattttat cctcatctca 2581 aacagtgtat atcattgtaa ccataaagag aaaggcagga tgatgaccag ggtgtagttg 2641 tttctaccaa taagaatatt tccacgccag ccagaattta tatgcagaaa tattctacct 2701 tatcatttaa ttataacaat tgttctctaa aactgtgctg aagtacaata taatataccc 2761 tgattgcctt gaaaaaaaag agattagaga aagtacttac aatctgacaa ataaacaaaa 2821 gtgaatttaa aaattcgtta caaatgcaag ctaaagttta acgaaaaagt tacagaaaat 2881 gaaaagaaaa taagaggaga caatggttgt caacagagta gaaagtgaaa gaaacaaaat 2941 tatcatgagg gtccatggtg atacaaggga catcttccca ttctaaacaa caccctgaaa 3001 actttgcccc ctccatataa catgaatttt acaatagcga aaaagaaaga acaatcaagg 3061 gtccccaaac tcaccctgaa gttctcagtc tagctcgacc tattcctttg ccctcggacg 3121 agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac 3181 ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat 3241 tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg 3301 atagagttgg tcaagaccaa tgcggagcat atacgcccgg agccgcggcg atcctgcaag 3361 ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct 3421 ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc 3481 aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga aatccgcgtg 3541 cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat cgagagcctg 3601 cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca catggggatc 3661 agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt gcggtccgaa 3721 tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca tggcctccgc 3781 gaccggctgc agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg 3841 tgcacggcgg gagatgcaat aggtcaggct ctcgctgaat tccccaatgt caagcacttc 3901 cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt tgtagaaacc 3961 atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga agctgaaagc 4021 acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga acttttcgat 4081 cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatggtgg cgactcgacg 4141 tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc ttttgaagcg 4201 tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc 4261 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa 4321 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca 4381 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct 4441 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac 4501 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc 4561 cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca gactgccttg 4621 ggaaaagcgc ctcccctacc cggtagactc gaccatagag cccaccgcat ccccagcatg 4681 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat 4741 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt 4801 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg 4861 gcaactagaa ggcacagctc gacttagatg gtgtcctggt cctgcgcgga tgtcaggtgg 4921 ggcggctgac ctgctgccgg ggtggatttc tttatcttct ggcccggaga agggcaggct 4981 gggatgtctg gggcctcggg ggtactgggg ctggaaacct cgtccaccga cagccgcagt 5041 ctcctgcggg ccggtggcct ggcctggtcg cgtccggggg cggcgggcgg gcgggcgcgc 5101 ttgcggctgc cactggtagc ggaggccggg gcgcggagtg cgggcctggc ggggctgggc 5161 tcgacatctg tagacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg 5221 cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg 5281 ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc 5341 tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg 5401 gcgccgtcct cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc 5461 attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg 5521 ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc 5581 acgcgctccc acttgaagcc ctcggggaag gacagcttca agtagtcggg aatgtcggcg 5641 gggtgcttca cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg 5701 aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg 5761 cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc 5821 accttgaagc gcatgaactc cttgatgatg gccatgttat cctcctcgcc cttgctcacc 5881 atggtggcga ctcgacgtcg aaaggcccgg agatgaggaa gaggagaaca gcgcggcaga 5941 cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag gaccttcggg cgcccgcccc 6001 gcccctgagc ccgcccctga gcccgccccc ggacccaccc cttcccagcc tctgagccca 6061 gaaagcgaag gagcaaagct gctattggcc gctgccccaa aggcctaccc gcttccattg 6121 ctcagcggtg ctgtccatct gcacgagact agtgagacgt gctacttcca tttgtcacgt 6181 cctgcacgac gcgagctgcg gggcgggggg gaacttcctg actaggggag gagtagaagg 6241 tggcgcgaag gggccaccaa agaacggagc cggttggcgc ctaccggtgg atgtggaatg 6301 tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc agcggggctg ctaaagcgca 6361 tgctccagac tgccttggga aaagcgcctc ccctacccgg tagactcgac gcctacccgc 6421 ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc tacttccatt 6481 tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac taggggagga 6541 gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct accggtggat 6601 gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag cggggctgct 6661 aaagcgcatg ctccagactg ccttgggaaa agcgcctccc ctacccggta gactcgacgc 6721 ctacccgctt ccattgctca gcggtgctgt ccatctgcac gagactagtg agacgtgcta 6781 cttccatttg tcacgtcctg cacgacgcga gctgcggggc gggggggaac ttcctgacta 6841 ggggaggagt agaaggtggc gcgaaggggc caccaaagaa cggagccggt tggcgcctac 6901 cggtggatgt ggaatgtgtg cgaggccaga ggccacttgt gtagcgccaa gtgcccagcg 6961 gggctgctaa agcgcatgct ccagactgcc ttgggaaaag cgcctcccct acccggtaga 7021 ctcgacgcct acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag 7081 acgtgctact tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt 7141 cctgactagg ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg 7201 gcgcctaccg gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt 7261 gcccagcggg gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac 7321 ccggtagact cgacgcctac ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga 7381 ctagtgagac gtgctacttc catttgtcac gtcctgcacg acgcgagctg cggggcgggg 7441 gggaacttcc tgactagggg aggagtagaa ggtggcgcga aggggccacc aaagaacgga 7501 gccggttggc gcctaccggt ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag 7561 cgccaagtgc ccagcggggc tgctaaagcg catgctccag actgccttgg gaaaagcgcc 7621 tcccctaccc ggtagactcg agttacgcgt aactcgagct ttggggcagc ggccaatagc 7681 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc 7741 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct 7801 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt 7861 cgacgtcgag tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 7921 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 7981 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 8041 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc 8101 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 8161 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 8221 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 8281 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg 8341 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 8401 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 8461 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag 8521 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 8581 gacgagctgt acaaggtcga gatgaatccc agtatgaagc agaaacaaga agaaatcaaa 8641 gagaatataa agaatagttc tgtcccaaga agaactctga agatgattca gccttctgca 8701 tctggatctc ttgttggaag agaaaatgag ctgtccgcag gcttgtccaa aaggaaacat 8761 cggaatgacc acttaacatc tacaacttcc agccctgggg ttattgtccc agaatctagt 8821 gaaaataaaa atcttggagg agtcacccag gagtcatttg atcttatgat taaagaaaat 8881 ccatcctctc agtattggaa ggaagtggca gaaaaacgga gaaaggcgct gtaagtcgag 8941 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 9001 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 9061 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 9121 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggtcgag c //