LOCUS       Exported               11897 bp ds-DNA     circular SYN 24-AUG-1994
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSGRNA-Pj23119
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11897)
  AUTHORS   Demo User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 27, 2017 from SnapGene 3.1.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11897
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1033..1368
                     /note="Rep(+)"
     CDS             complement(1966..2592)
                     /note="Tet R"
     misc_feature    complement(2722..3294)
                     /note="SecY-F"
     D_segment       3309..3445
                     /note="sgRNA"
     promoter        3309..3343
                     /note="pJ23119"
     CDS             3344..3363
                     /note="N20"
     CDS             4134..4784
                     /note="cat R"
     promoter        4964..5060
                     /note="Pxyl/tet-RBS"
     CDS             5061..9167
                     /note="cas9"
     misc_feature    complement(9212..9228)
                     /note="M13-R Primer"
     misc_binding    complement(9234..9256)
                     /note="LacO"
     rep_origin      complement(9650..10278)
                     /direction=LEFT
                     /note="ColE1 origin"
     CDS             10430..11089
                     /note="Amp R"
     CDS             complement(11726..11794)
                     /note="LacZ alpha Reporter"
ORIGIN
        1 ctgcagaacg gattgttgat gattacgaaa atattaagag cacagactat tacacagaaa
       61 atcaagaatt aaaaaaacgt agagagagtt tgaaagaagt agtgaataca tggaaagagg
      121 ggtatcacga aaaaagtaaa gaggttaata aattaaagcg agagaatgat agtttgaatg
      181 agcagttgaa tgtatcagag aaatttcaag ctagtacagt gactttatat cgtgctgcga
      241 gggcgaattt ccctgggttt gagaaagggt ttaataggct taaagagaaa ttctttaatg
      301 attccaaatt tgagcgtgtg ggacagttta tggatgttgt acaggataat gtccagaagg
      361 tcgatagaaa gcgtgagaaa cagcgtacag acgatttaga gatgtagagg tacttttatg
      421 ccgagaaaac tttttgcgtg tgacagtcct taaaatatac ttagagcgta agcgaaagta
      481 gtagcgacag ctattaactt tcggttgcaa agctctagga tttttaatgg acgcagcgca
      541 tcacacgcaa aaaggaaatt ggaataaatg cgaaatttga gatgttaatt aaagaccttt
      601 ttgaggtctt tttttcttag atttttgggg ttatttaggg gagaaaacat aggggggtac
      661 tacgacctcc cccctaggtg tccattgtcc attgtccaaa caaataaata aatattgggt
      721 ttttaatgtt aaaaggttgt tttttatgtt aaagtgaaaa aaacagatgt tgggaggtac
      781 agtgatagtt gtagatagaa aagaagagaa aaaagttgct gttactttaa gacttacaac
      841 agaagaaaat gagatattaa atagaatcaa agaaaaatat aatattagca aatcagatgc
      901 aaccggtatt ctaataaaaa aatatgcaaa ggaggaatac ggtgcatttt aaacaaaaaa
      961 agatagacag cactggcatg ctgcctatct atgactaaat tttgttaaat gtattagcac
     1021 cgttattata tcatgagcga aaatgtaata aaagaaactg aaaacaagaa aaattcaaga
     1081 ggacgtaatt ggacatttgt tttatatcca gaatcagcaa aagccgagtg gttagagtat
     1141 ttaaaagagt tacacattca atttgtagtg tctccattac atgataggga tactgataca
     1201 gaagatagga tgaaaaaaga gcattatcat attctagtga tgtatgaggg taataaatct
     1261 tatgaacaga taaaaataat taacagaaga attgaatgcg actattccgc agattgcagg
     1321 aagtgtgaaa ggtcttgtga gatatatgct tcacatggac gatcctaata aatttaaata
     1381 tcaaaaagaa gatatgatag tttatggcgg tgtagatgtt gatgaattat taaagaaaac
     1441 aacaacagat agatataaat taattaaaga aatgattgag tttattgatg aacaaggaat
     1501 cgtagaattt aagagtttaa tggattatgc aatgaagttt aaatttgatg attggttccc
     1561 gcttttatgt gataactcgg cgtatgttat tcaagaatat ataaaatcaa atcggtataa
     1621 atctgaccga tagattttga atttaagagt gtcacaagac actctttttt cgcaccagcg
     1681 aaaactggtt taagccgact gcgcaaaaga cataatcgat tcacaaaaaa taggcacacg
     1741 aaaaacaagt taagggatgc agtttatgca tcccttaact tacttattaa ataatttata
     1801 gctattgaaa agagataaga attgttcaaa gctaatattg tttaaatcgt caattcctgc
     1861 atgttttaag gaattgttaa attgattttt tgtaaatatt ttcttgtatt ctttgttatc
     1921 ttggttaccg tgaagttacc atcacggaaa aaggttatgc tgcttttaag acccactttc
     1981 acatttaagt tgtttttcta atccgcatat gatcaattca aggccgaata agaaggctgg
     2041 ctctgcacct tggtgatcaa ataattcgat agcttgtcgt aataatggcg gcatactatc
     2101 agtagtaggt gtttcccttt cttctttagc gacttgatgc tcttgatctt ccaatacgca
     2161 acctaaagta aaatgcccca cagcgctgag tgcatataat gcattctcta gtgaaaaacc
     2221 ttgttggcat aaaaaggcta attgattttc gagagtttca tactgttttt ctgtaggccg
     2281 tgtacctaaa tgtacttttg ctccatcgcg atgacttagt aaagcacatc taaaactttt
     2341 agcgttatta cgtaaaaaat cttgccagct ttccccttct aaagggcaaa agtgagtatg
     2401 gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat tttttacatg
     2461 ccaatacaat gtaggctgct ctacacctag cttctgggcg agtttacggg ttgttaaacc
     2521 ttcgattccg acctcattaa gcagctctaa tgcgctgtta atcactttac ttttatctaa
     2581 tctagacatc attaattcct cctttttgtt gacactctat cattgataga gttatttgtc
     2641 aaactagttt tttatttgga tcccctcgag ttcatgaaaa actaaaaaaa atattgacac
     2701 tctatcattg atagagtata attaaaataa gcttgatggg aatatgataa tagaaatacc
     2761 attaccaaca ccgaactgag tgatttgatc accaagccat attaagaaag cagttcctgc
     2821 tgtcaaaact agtgctatta ataaataact cataattgac tgattgataa tcagcgcacc
     2881 tttgagataa ttattaaatt ggaatgccat acctatagat tggataaatg ctaaagaaat
     2941 tgctaaataa cgagtaacgt tatttaactt tcttctacct acttcacctt gttttgccca
     3001 ttctgagaat ttagggacaa tatccatttg taataattgc attacgattg atgcagtgat
     3061 gtagggtaca atacccattg caaaaataga aaatcgtttc aaggctccgc caccaaaagt
     3121 atttaataac tcagtggcac cttgagaacc ttggggatta tcaaaagctg caggatttac
     3181 tcctggagct ggtatataag tccctatttt aaaaattact aacattgcta gtgtgaagaa
     3241 aatcttgtta cgaacctctt ttgttctaaa gaagttcaca agggtttgaa tcattagatc
     3301 acagatcttt gacagctagc tcagtcctag gtataatact agtatgtgtt cgtatgttac
     3361 tgcgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     3421 agtggcaccg agtcggtgct ttttttgcgg ccgcgtgatc taatgattca tatctacatc
     3481 aaatgctggg gcatatgttt taccctcagg tacaggtaag tcacccgctt caaacgtctt
     3541 cttaatatgc gtaattgcat ctactctaaa tccatcaatg cctttatcaa accaccagtt
     3601 catcatttca aatacagcat ctctaacttc cggattaccc caattcaaat caggttgttt
     3661 tttactgaat aaatggaaat aatattgctc agtattagca tcatattccc gcgcgccaat
     3721 tgagctcccc tttctatgta tgttttttac tagtcattta aaacgataca ttaataggta
     3781 cgaaaaagca actttttttg cgcttaaaac cagtcatacc aataacttaa gggtaactag
     3841 cctcgccggc aatagttacc cttattatca agataagaaa gaaaaggatt tttcgctacg
     3901 ctcaaatcct ttaaaaaaac acaaaagacc acatttttta atgtggtctt ttattcttca
     3961 actaaagcac ccattagttc aacaaacgaa aattggataa agtgggatat ttttaaaata
     4021 tatatttatg ttacagtaat attgactttt aaaaaaggat tgattctaat gaagaaagca
     4081 gacaagtaag cctcctaaat tcactttaga taaaaattta ggaggcatat caaatgaaat
     4141 ttaataaaat tgatttagac aattggaaga gaaaagagat atttaatcat tatttgaacc
     4201 aacaaacgac ttttagtata accacagaaa ttgatattag tgttttatac cgaaacataa
     4261 aacaagaagg atataaattt taccctgcat ttattttctt agtgacaagg gtgataaact
     4321 caaatacagc ttttagaact ggttacaata gcgacggaga gttaggttat tgggataagt
     4381 tagagccact ttatacaatt tttgatggtg tatctaaaac attctctggt atttggactc
     4441 ctgtaaagaa tgacttcaaa gagttttatg atttatacct ttctgatgta gagaaatata
     4501 atggttcggg gaaattgttt cccaaaacac ctatacctga aaatgctttt tctctttcta
     4561 ttattccatg gacttcattt actgggttta acttaaatat caataataat agtaattacc
     4621 ttctacccat tattacagca ggaaaattca ttaataaagg taattcaata tatttaccgc
     4681 tatctttaca ggtacatcat tctgtttgtg atggttatca tgcaggattg tttatgaact
     4741 ctattcagga attgtcagat aggcctaatg actggctttt ataatatgag ataatgccga
     4801 ctgtactttt tacagtcggt tttctaatgt cactaacctg ccccgttagt tgaagaaggt
     4861 ttttatatta cagctccaga tccatatcct tctttttctg aaccgacttc tcctttttcg
     4921 cttctttatt ccaattgctt tattgacgtt gagcctcgga accgtcgact ttatttggat
     4981 cccctcgagt tcatgaaaaa ctaaaaaaaa tattgacact ctatcattga tagagtataa
     5041 ttaacaagga ggaataaaaa atggataaga aatactcaat aggcttagat atcggcacaa
     5101 atagcgtcgg atgggcggtg atcactgatg attataaggt tccgtctaaa aagttcaagg
     5161 ttctgggaaa tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg
     5221 acagtggaga gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac
     5281 gtcggaagaa tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag
     5341 atgatagttt ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg
     5401 aacgtcatcc tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa
     5461 ctatctatca tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa
     5521 tctatttggc cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt
     5581 taaatcctga taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc
     5641 aattatttga agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg
     5701 cacgattgag taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga
     5761 aaaatggctt atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat
     5821 caaattttga tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg
     5881 atttagataa tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta
     5941 agaatttatc agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta
     6001 aggctcccct atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc
     6061 ttttaaaagc tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc
     6121 aatcaaaaaa cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata
     6181 aatttatcaa accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa
     6241 atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa
     6301 ttcacttggg tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa
     6361 aagacaatcg tgagaagatt gaaaaaatct tgacttttcg aattccttat tatgttggtc
     6421 cattggcgcg tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta
     6481 ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac
     6541 gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc
     6601 tttatgagta ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa
     6661 tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca
     6721 aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat
     6781 gttttgatag tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct
     6841 accatgattt gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag
     6901 atatcttaga ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg
     6961 aaagacttaa aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc
     7021 gccgttatac tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc
     7081 aatctggcaa aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta
     7141 tgcagctgat ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt
     7201 ctggacaagg cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta
     7261 aaaaaggtat tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc
     7321 ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc
     7381 agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc
     7441 agattcttaa agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct
     7501 attatctcca aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa
     7561 gtgattatga tgtcgatcac attgttccac aaagtttcct taaagacgat tcaatagaca
     7621 ataaggtctt aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag
     7681 aagtagtcaa aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc
     7741 aacgtaagtt tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag
     7801 ctggttttat caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa
     7861 ttttggatag tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta
     7921 aagtgattac cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata
     7981 aagtacgtga gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg
     8041 gaactgcttt gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata
     8101 aagtttatga tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg
     8161 caaaatattt cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa
     8221 atggagagat tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct
     8281 gggataaagg gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata
     8341 ttgtcaagaa aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa
     8401 gaaattcgga caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt
     8461 ttgatagtcc aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat
     8521 cgaagaagtt aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct
     8581 ttgaaaaaaa tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact
     8641 taatcattaa actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc
     8701 tggctagtgc cggagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga
     8761 attttttata tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac
     8821 aaaaacaatt gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca
     8881 gtgaattttc taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat
     8941 ataacaaaca tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta
     9001 cgttgacgaa tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta
     9061 aacgatatac gtctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg
     9121 gtctttatga aacacgcatt gatttgagtc agctaggagg tgactgaagt atattttaga
     9181 tgaagattat ttcttaagaa ttcgtgacta tgtcatagct gtttcctgtg tgaaattgtt
     9241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgaaag cctggggtgc
     9301 ctaaaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     9361 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     9421 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     9481 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     9541 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     9601 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     9661 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     9721 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     9781 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     9841 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     9901 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     9961 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    10021 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    10081 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
    10141 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    10201 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    10261 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
    10321 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
    10381 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
    10441 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
    10501 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    10561 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    10621 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    10681 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    10741 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    10801 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    10861 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    10921 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    10981 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    11041 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    11101 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    11161 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    11221 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    11281 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    11341 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    11401 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
    11461 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
    11521 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
    11581 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
    11641 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
    11701 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
    11761 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
    11821 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
    11881 gtgccaagct tgcatgc
//
