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
//
