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
//
