LOCUS       Exported                6423 bp ds-DNA     circular SYN 01-JUN-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCas56 YFP sgRNA8
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 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..6423
                     /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"
     misc_feature    1337..1377
                     /note="T7 terminator"
     gene            complement(1413..1449)
                     /note="ssRa"
     gene            complement(1450..2069)
                     /note="tetR"
     RBS             2076..2087
                     /note="RBS"
     misc_feature    2096..2130
                     /note="J23113 Promoter"
     misc_feature    2132..2205
                     /note="Promoter"
     misc_feature    2152..2168
                     /note="TetO1"
     -10_signal      2169..2174
                     /note="-35"
     misc_feature    2175..2191
                     /note="TetO1"
     -10_signal      2192..2198
                     /note="-10"
     misc_feature    2199
                     /note="TSS from EKZ"
     misc_feature    2206..2225
                     /note="sgRNA (+172)"
     misc_feature    2226..2267
                     /note="cas9 Handle"
     misc_feature    2268..2308
                     /note="S. pyogenes terminator"
     misc_feature    2309..2409
                     /note="Spacer Included in Qi et. al. 2013"
                     /note="Spacer Included in Qi et. al. 2013 ""Repurposing 
                     CRISPR..."""
     terminator      2410..2675
                     /note="rrnB_terminator"
     misc_feature    2676..3676
                     /note="KanR Cassette"
     CDS             2777..3592
                     /note="KanR"
     misc_feature    3678..4188
                     /note="NS1 Bottom"
     gene            complement(3768..4160)
                     /note="SYNPCC7002_A0936"
     misc_feature    complement(3871..3895)
                     /note="152"
     misc_feature    complement(3881..3902)
                     /note="GG192 used in paper prove seg reverse"
     misc_feature    4189..4213
                     /note="Untitled"
     rep_origin      complement(4604..5192)
                     /direction=LEFT
                     /note="pMB1 origin of replication"
                     /note="pMB1 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     misc_RNA        complement(4604..5156)
                     /note="RNAII transcript (complementary strand)"
     -35_signal      5010..5015
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      5032..5037
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        5046..5153
                     /note="RNAI transcript"
     -10_signal      complement(5166..5171)
                     /note="RNAII promoter"
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(5187..5192)
                     /note="RNAII promoter"
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(5363..6223)
                     /codon_start=1
                     /note="beta-lactamase"
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     -10_signal      complement(6267..6272)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(6288..6293)
                     /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 caataactat aaacgcagaa aggcccaccc gaaggtgagc cagtgtgact
     1321 ctagtagaga gcgttcaacc ccttggggcc tctaaacggg tcttgagggg ttttttgtct
     1381 agtagtgatc tacactagca ctatcagtgt tattaagcta ctaaagcgta gttttcgtcg
     1441 tttgcagcgg acccactttc acatttaagt tgtttttcta atccgcatat gatcaattca
     1501 aggccgaata agaaggctgg ctctgcacct tggtgatcaa ataattcgat agcttgtcgt
     1561 aataatggcg gcatactatc agtagtaggt gtttcccttt cttctttagc gacttgatgc
     1621 tcttgatctt ccaatacgca acctaaagta aaatgcccca cagcgctgag tgcatataat
     1681 gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta attgattttc gagagtttca
     1741 tactgttttt ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg atgacttagt
     1801 aaagcacatc taaaactttt agcgttatta cgtaaaaaat cttgccagct ttccccttct
     1861 aaagggcaaa agtgagtatg gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa
     1921 gcccgcttat tttttacatg ccaatacaat gtaggctgct ctacacctag cttctgggcg
     1981 agtttacggg ttgttaaacc ttcgattccg acctcattaa gcagctctaa tgcgctgtta
     2041 atcactttac ttttatctaa tctggacatc tagtatttct cctctttctc tagtagctag
     2101 cataatccct aggactgagc tagccatcag gttaacaaaa aagcaggaat atccctatca
     2161 gtgatagaca gacatcccta tcagtgatag agtataaagt taactcgtcg ccgtccagct
     2221 cgaccgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     2281 aaagtggcac cgagtcggtg ctttttttga agcttgggcc cgaacaaaaa ctcatctcag
     2341 aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct
     2401 ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     2461 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     2521 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     2581 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2641 gggcctttcg ttttatctgt tgtttgtcgg tgaactacgg ggtctgacgc tcagtggaac
     2701 gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag
     2761 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt
     2821 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2881 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2941 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     3001 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3061 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     3121 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3181 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3241 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3301 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3361 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3421 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3481 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3541 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat
     3601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3661 catttccccg aaaaggtgga tccgaaaata aagtcaaagt gcttctataa tttaagatct
     3721 atttttaaac agtacagctt tacagaaaga aaaaccagaa gggacgccta ttttatctcg
     3781 ttttcttccc agagtagcag gacatttttt tctgtaaaag cagagcaatt tgtactgaaa
     3841 tcattatcaa taatgacatt agattcttcg atcaagacat ctgtaccgtt gcttatatct
     3901 gcttcagagc ctaatacaat ggggtattgt ggttgttgac atgcagcatt aacgagtact
     3961 ttgactggga atacaaccga atcatcgcta taggtgtaga cattggcgtg gtagtaatag
     4021 gtatagtcat catagtcgta attatattca taaccatacc gatactggta atcaaaatca
     4081 ataaatacag gttcactttt tgcctgttga acaacgataa atgatagacc actaagggct
     4141 gctgtaaaca gtttattcat tttgctttct ccatcccttg acttaatttg gcgtaatcat
     4201 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4261 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4321 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4381 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6301 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6361 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6421 gtc
//
