LOCUS Exported 5538 bp ds-DNA circular SYN 21-MAR-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCas28 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5538) AUTHORS . TITLE Direct Submission JOURNAL Exported Monday, Mar 21, 2016 from SnapGene 3.1.1 http://www.snapgene.com COMMENT LOCUS pUC19 2686 bp DNA circular 18-OCT-2007 DEFINITION Cloning vector pUC19, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pUC19 ORGANISM Cloning vector pUC19 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 2686) AUTHORS New England Biolabs. TITLE Direct Submission JOURNAL Submitted (18-OCT-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA COMMENT See also GenBank accession L09137. FEATURES Location/Qualifiers source 1..5538 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(146..397) /codon_start=1 /note="beta-galactosidase alpha fragment" /translation="SLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQ LRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA" misc_feature 397..1276 /note="NS1 Top" gene 397..1014 /note="SYNPCC7002_A0935" promoter 1285..1319 /note="pJ23108" /note="pJ23119" misc_feature 1321..1340 /note="20 bp Specificity Determining Region" misc_feature 1341..1382 /note="cas9 Handle" misc_feature 1383..1423 /note="S. pyogenes terminator" misc_feature 1424..1524 /note="Spacer Included in Qi et. al. 2013" /note="Spacer Included in Qi et. al. 2013 ""Repurposing CRISPR...""" terminator 1525..1790 /note="rrnB_terminator" misc_feature 1791..2791 /note="KanR Cassette" CDS 1892..2707 /note="KanR" misc_feature 2793..3303 /note="NS1 Bottom" gene complement(2883..3275) /note="SYNPCC7002_A0936" rep_origin complement(3719..4307) /direction=LEFT /note="pMB1 origin of replication" /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" misc_RNA complement(3719..4271) /note="RNAII transcript (complementary strand)" -35_signal 4125..4130 /note="RNAI promoter (clockwise); TTGAAG" -10_signal 4147..4152 /note="RNAI promoter (clockwise); GCTACA" misc_RNA 4161..4268 /note="RNAI transcript" -10_signal complement(4281..4286) /note="RNAII promoter" /note="RNAII promoter (counter-clockwise); CGTAAT" -35_signal complement(4302..4307) /note="RNAII promoter" /note="RNAII promoter (counter-clockwise); TTGAGA" CDS complement(4478..5338) /codon_start=1 /note="beta-lactamase" /note="ampR (confers resistance to ampicillin)" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" -10_signal complement(5382..5387) /note="bla promoter (counter-clockwise); GAGACA" -35_signal complement(5403..5408) /note="bla promoter (counter-clockwise); TTCAAA" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgatgt ttggtcgaaa attcttcaca 421 cttttaacgg ctgtctccac agtgttcctg actcctattt acggagcggc tcaaacgatg 481 caagatcaaa gtaatacgat gaataatgaa aattcattgg ggaatagtgc gactggggta 541 gtcaaccaaa ccgctgtgta caacttaggt caaagtgcca cctacggttt tagagggatc 601 gtttgtcccc gcccttcctt tgtgctcagt ggctctgcaa atggcaatac atttgatggg 661 gagagttcaa attcctacgc tctttccggt gccattattg tgcccctcgg tggtcaagtg 721 ggcagcaact gtagagattt cagcactacc ctactggaaa gagaacaagc cgaattagca 781 aagtttcaaa tccaatcttc agcggaaaca gtcaaacttt gtgctgacat gatttcttca 841 ggcattcaag tagatccaga ggtttatcca gtactggcgc ggcaatgttc tgcagttcgg 901 ttggtttcac aaggacgggc ttctagcaca aatgaagccc gacaaaatga atcgactcac 961 aataagctag atgatgatgt agaattgtct gctaattact taaaaactga gtaaatataa 1021 tattttgaat agtgaggaaa aatatctcaa aggctatatg aaaataaaaa aagagcggct 1081 aaaaaatctt cagcttacag tgacgcctgt ctttgcattc taaagcacta aaacacagtc 1141 gtttttttac tgactccaag tgtttttatt actatactta gaaattcgtc aaataggcat 1201 gaaattgctg tattaaaata tcctactgta tcttttagtt cacactattt ttcttgagtt 1261 aattttgtat caataacgga attcctgaca gctagctcag tcctaggtat aatgctagct 1321 ggcatcgccc tcgccctcgc gttttagagc tagaaatagc aagttaaaat aaggctagtc 1381 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgaagctt gggcccgaac 1441 aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt 1501 gagtttaaac ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat 1561 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag 1621 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg 1681 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg 1741 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac tacggggtct 1801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc 1861 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg 1921 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg 1981 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc 2041 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt 2101 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac 2161 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt 2221 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg 2281 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc 2341 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg 2401 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc 2461 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa 2521 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc 2581 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa 2641 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt 2701 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 2761 taggggttcc gcgcacattt ccccgaaaag gtggatccga aaataaagtc aaagtgcttc 2821 tataatttaa gatctatttt taaacagtac agctttacag aaagaaaaac cagaagggac 2881 gcctatttta tctcgttttc ttcccagagt agcaggacat ttttttctgt aaaagcagag 2941 caatttgtac tgaaatcatt atcaataatg acattagatt cttcgatcaa gacatctgta 3001 ccgttgctta tatctgcttc agagcctaat acaatggggt attgtggttg ttgacatgca 3061 gcattaacga gtactttgac tgggaataca accgaatcat cgctataggt gtagacattg 3121 gcgtggtagt aataggtata gtcatcatag tcgtaattat attcataacc ataccgatac 3181 tggtaatcaa aatcaataaa tacaggttca ctttttgcct gttgaacaac gataaatgat 3241 agaccactaa gggctgctgt aaacagttta ttcattttgc tttctccatc ccttgactta 3301 atttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 3361 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 3421 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 3481 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 3541 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 3601 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 3661 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 3721 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 3781 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 3841 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 3901 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 3961 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4021 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 4081 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 4141 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 4201 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 4261 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4321 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4381 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4441 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 4501 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4561 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 4621 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 4681 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 4741 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 4801 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 4861 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 4921 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 4981 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5041 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5101 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5161 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5221 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5281 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5401 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 5461 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 5521 tcacgaggcc ctttcgtc //