LOCUS Exported 5536 bp ds-DNA circular SYN 24-MAR-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCas15 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5536) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Mar 24, 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..5536 /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="pJ23119" misc_feature 1321..1338 /note="18 bp Specificity Determining Region" /note="20 bp Specificity Determining Region" misc_feature 1339..1380 /note="cas9 Handle" misc_feature 1381..1421 /note="S. pyogenes terminator" misc_feature 1422..1522 /note="Spacer Included in Qi et. al. 2013" /note="Spacer Included in Qi et. al. 2013 ""Repurposing CRISPR...""" terminator 1523..1788 /note="rrnB_terminator" misc_feature 1789..2789 /note="KanR Cassette" CDS 1890..2705 /note="KanR" misc_feature 2791..3301 /note="NS1 Bottom" gene complement(2881..3273) /note="SYNPCC7002_A0936" rep_origin complement(3717..4305) /direction=LEFT /note="pMB1 origin of replication" /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" misc_RNA complement(3717..4269) /note="RNAII transcript (complementary strand)" -35_signal 4123..4128 /note="RNAI promoter (clockwise); TTGAAG" -10_signal 4145..4150 /note="RNAI promoter (clockwise); GCTACA" misc_RNA 4159..4266 /note="RNAI transcript" -10_signal complement(4279..4284) /note="RNAII promoter" /note="RNAII promoter (counter-clockwise); CGTAAT" -35_signal complement(4300..4305) /note="RNAII promoter" /note="RNAII promoter (counter-clockwise); TTGAGA" CDS complement(4476..5336) /codon_start=1 /note="beta-lactamase" /note="ampR (confers resistance to ampicillin)" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" -10_signal complement(5380..5385) /note="bla promoter (counter-clockwise); GAGACA" -35_signal complement(5401..5406) /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 attcttgaca gctagctcag tcctaggtat aatgctagct 1321 catcgccctc gccctcgcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg 1381 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgaagcttgg gcccgaacaa 1441 aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga 1501 gtttaaacgg tctccagctt ggctgttttg gcggatgaga gaagattttc agcctgatac 1561 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg 1621 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta 1681 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct 1741 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacta cggggtctga 1801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg 1861 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct 1921 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg 1981 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag 2041 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca 2101 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc 2161 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag 2221 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt 2281 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc 2341 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta 2401 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg 2461 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat 2521 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca 2581 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat 2641 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt 2701 tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 2761 ggggttccgc gcacatttcc ccgaaaaggt ggatccgaaa ataaagtcaa agtgcttcta 2821 taatttaaga tctattttta aacagtacag ctttacagaa agaaaaacca gaagggacgc 2881 ctattttatc tcgttttctt cccagagtag caggacattt ttttctgtaa aagcagagca 2941 atttgtactg aaatcattat caataatgac attagattct tcgatcaaga catctgtacc 3001 gttgcttata tctgcttcag agcctaatac aatggggtat tgtggttgtt gacatgcagc 3061 attaacgagt actttgactg ggaatacaac cgaatcatcg ctataggtgt agacattggc 3121 gtggtagtaa taggtatagt catcatagtc gtaattatat tcataaccat accgatactg 3181 gtaatcaaaa tcaataaata caggttcact ttttgcctgt tgaacaacga taaatgatag 3241 accactaagg gctgctgtaa acagtttatt cattttgctt tctccatccc ttgacttaat 3301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 3361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 3421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 3481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 3541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 3601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 3661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 3721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 3781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 3841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 3901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 3961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 4021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 4081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 4141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 4201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 4261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 4321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 4381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 4441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 4501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 4561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 4621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 4681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 4741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 4801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 4861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 4921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 4981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 5041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 5101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 5161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 5221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 5281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 5341 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 5401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 5461 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc 5521 acgaggccct ttcgtc //