LOCUS       Exported                5545 bp ds-DNA     circular SYN 01-MAY-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCas45_sgRNA YFP pJ23119 7 extra 5'
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, May 1, 2016 from SnapGene 3.1.2
            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..5545
                     /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..1327
                     /note="extra from Tet promote constructs"
                     /note="Promoter"
     misc_feature    1321
                     /note="TSS from EKZ"
     misc_feature    1328..1347
                     /note="20 bp Specificity Determining Region"
     misc_feature    1348..1389
                     /note="cas9 Handle"
     misc_feature    1390..1430
                     /note="S. pyogenes terminator"
     misc_feature    1431..1531
                     /note="Spacer Included in Qi et. al. 2013"
                     /note="Spacer Included in Qi et. al. 2013 ""Repurposing 
                     CRISPR..."""
     terminator      1532..1797
                     /note="rrnB_terminator"
     misc_feature    1798..2798
                     /note="KanR Cassette"
     CDS             1899..2714
                     /note="KanR"
     misc_feature    2800..3310
                     /note="NS1 Bottom"
     gene            complement(2890..3282)
                     /note="SYNPCC7002_A0936"
     CDS             complement(2890..3282)
                     /codon_start=1
                     /note="hypothetical protein"
                     /translation="MNKLFTAALSGLSFIVVQQAKSEPVFIDFDYQYRYGYEYNYDYDD
                     YTYYYHANVYTYSDDSVVFPVKVLVNAACQQPQYPIVLGSEADISNGTDVLIEESNVII
                     DNDFSTNCSAFTEKNVLLLWEENEIK"
     protein_bind    complement(3422..3434)
                     /note="CAP protein binding site"
     rep_origin      complement(3726..4314)
                     /direction=LEFT
                     /note="pMB1 origin of replication"
                     /note="pMB1 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     misc_RNA        complement(3726..4278)
                     /note="RNAII transcript (complementary strand)"
     -35_signal      4132..4137
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      4154..4159
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        4168..4275
                     /note="RNAI transcript"
     -10_signal      complement(4288..4293)
                     /note="RNAII promoter"
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(4309..4314)
                     /note="RNAII promoter"
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(4485..5345)
                     /codon_start=1
                     /note="beta-lactamase"
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     -10_signal      complement(5389..5394)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(5410..5415)
                     /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 gttaactggc atcgccctcg ccctcgcgtt ttagagctag aaatagcaag ttaaaataag
     1381 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gaagcttggg
     1441 cccgaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat
     1501 catcattgag tttaaacggt ctccagcttg gctgttttgg cggatgagag aagattttca
     1561 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1621 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1681 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1741 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaactac
     1801 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa
     1861 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     1921 cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1981 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     2041 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     2101 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     2161 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     2221 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2281 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2341 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2401 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     2461 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2521 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2581 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2641 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2701 atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2761 aaacaaatag gggttccgcg cacatttccc cgaaaaggtg gatccgaaaa taaagtcaaa
     2821 gtgcttctat aatttaagat ctatttttaa acagtacagc tttacagaaa gaaaaaccag
     2881 aagggacgcc tattttatct cgttttcttc ccagagtagc aggacatttt tttctgtaaa
     2941 agcagagcaa tttgtactga aatcattatc aataatgaca ttagattctt cgatcaagac
     3001 atctgtaccg ttgcttatat ctgcttcaga gcctaataca atggggtatt gtggttgttg
     3061 acatgcagca ttaacgagta ctttgactgg gaatacaacc gaatcatcgc tataggtgta
     3121 gacattggcg tggtagtaat aggtatagtc atcatagtcg taattatatt cataaccata
     3181 ccgatactgg taatcaaaat caataaatac aggttcactt tttgcctgtt gaacaacgat
     3241 aaatgataga ccactaaggg ctgctgtaaa cagtttattc attttgcttt ctccatccct
     3301 tgacttaatt tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3361 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3421 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3481 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3541 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3601 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3661 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3721 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3781 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3841 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3901 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3961 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4021 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4081 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4141 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4201 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4261 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4321 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4381 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4441 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4501 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4561 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4621 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4681 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4741 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4801 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4861 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4921 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4981 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5041 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5101 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5161 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5221 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5281 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5341 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5401 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5461 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5521 aggcgtatca cgaggccctt tcgtc
//
