LOCUS       Exported               11077 bp ds-DNA     circular SYN 15-Ò»ÔÂ-2016
DEFINITION  synthetic circular DNA
ACCESSION   pIMP1
VERSION     .
KEYWORDS    pNICKclos1.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11077)
  AUTHORS   Self
  JOURNAL   Unpublished.
REFERENCE   2  (bases 1 to 11077)
  AUTHORS   rc
  TITLE     Direct Submission
  JOURNAL   Exported 2016Äê1ÔÂ28ÈÕ from SnapGene Viewer 2.8.2
            http://www.snapgene.com
COMMENT     SECID/File created by SciEd Central, Scientific & Educational 
            Software
            ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..11077
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..150
                     /note="sgRNA-cac0027"
     misc_feature    14..48
                     /note="Pj23119"
     misc_feature    49..68
                     /note="20bp-spacer"
     misc_feature    69..150
                     /note="tracrRNA"
     misc_feature    159..1158
                     /note="cac0027-HA-up"
     misc_feature    1165..2164
                     /note="cac0027-HA-dn"
     misc_feature    2177..2292
                     /note="ptb promotor"
     misc_feature    2281..2286
                     /note="RBS"
     misc_feature    2299..6405
                     /note="cas9-nickase"
     misc_feature    6724..6747
                     /note="3' sequencing:primer"
     CDS             7012..7869
                     /codon_start=1
                     /gene="bla"
                     /product="B-lactamase"
                     /note="bla"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    8630..8692
                     /note="ori"
     CDS             9401..10132
                     /codon_start=1
                     /gene="mslR"
                     /note="mslR"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVQRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIFGNIPY
                     NISTDIIRKIVFDSIADEIYLIVEYGFAKRLLNTKRSLALFLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLNRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     rep_origin      10583..11020
                     /direction=RIGHT
                     /gene="repL"
                     /note="repL"
     misc_feature    11018..11041
                     /note="5' sequencing:primer"
ORIGIN
        1 agcttggctg cagttgacag ctagctcagt cctaggtata atactagtta tacttaaatc
       61 acaggctggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      121 gaaaaagtgg caccgagtcg gtgctttttt gcggccgcat tcttatgaaa attcaagtgc
      181 tctaactgta atttttccag gaataaatta tggtcatgaa aagcctcttc tgtattatgc
      241 aaggaaagca gctgcacaga ataatataga tattctatgc atcaactata agaataagat
      301 ttcttgggag gacataggaa aacaaagtat tgaagatgta gctattgaaa taaagggcat
      361 tataaaaaat gttattgaaa aaaagtatga taacatttat tttttaggga aaagtgttgg
      421 aacagaaata gctggatgta catcttctaa attaaatata gataatataa agtttcttta
      481 cttaacacct attgaagaga ctttaaagta tatagttgac agaaagtgca ttgtggtgtc
      541 aggcactaag gatgaatttt ttaaaagtga atgtataaat aaaatcagag aaaataaaaa
      601 tgttaaattg atgttgtttg agggagctaa tcattctctt gagattaagg atgatattat
      661 aagaagttta agaattttag ataacgtagc acaattgtat ttggacttct ttaaataaaa
      721 acatggaggg ctaaaaatgg aacaatacaa acaagaattt atagaattta tggtggagag
      781 taatgtactt acctttgggg atttcataac taaaagcggc agaagaacac cattttttat
      841 aaatacaggt aactacaaga caggtaatca attaaataag ttggctaagt tttatgctaa
      901 agcaatatat gataattttg gagatgattt tgatatttta tttgggcctg catataaagg
      961 aataccttta agtgtttcag tagctatggc acttgataat atttatggaa ttaatgcagc
     1021 ttattgttca aatagaaaag aagttaaaga tcacggtgat aagggaatac ttcttggagc
     1081 aaagcttgaa gaaggagaca gagttataat tgtagaagat gtcacaacag ctggtacatc
     1141 agtatacgaa acaatgccga tatcaggttg atgtaaaggg aatcataata tcagtggata
     1201 gaatggaaag aggtaaggga gataagagtg ccttaactga acttaaagaa aagtttggat
     1261 ttaaaacatg ttctattgtt actatggaag aggtagtaga atatttgtat aagaaaaata
     1321 tcaatggcaa agtaatcata gatgataaaa tgaaagatag aattaatgag tactataaag
     1381 agtatggagt aaaatagtaa aataaatgtg cctcaactta gatgttaagg cacatttatt
     1441 ttatatatta ttcatgtttt gaaacatttt tatcttttgt gtattttacg tgtagtaatt
     1501 tgtgagcaag tccttcacct ggttttccaa agtagctatc atacatttta ataatagctg
     1561 gattatcatg tgactttctc tttgaaagaa catttttatc ttggttgtat aatactgatg
     1621 ctcttagttt tctgtaatca acattttctc tatcaagagc atttacgtga ggttgacctc
     1681 caccatttat acatccacca gggcaagcca ttacttctat aaagtgatat tgtttttcgt
     1741 tcatttttcc agatttcata aactcgaaga agttagaagc accatttata acagcaacgt
     1801 ttagtttatt tccagcaatt tcaacttccg cttcttttat gcctttaaag cctcttactt
     1861 cagtgtaatc aacattttca agttctttat tttcagcaaa gtctttagct gatcttattg
     1921 cagcttccat aacgccaccg gttgcaccaa agatagctcc agcaccactg taagtaccca
     1981 tagcaggatc aacttcacca tcttcaagat ctgcaaattt aatttttgca tctttaatca
     2041 tttttgcaag ctctcttgta gttaaggatg catcaatatc tcttaagctg ttagtttcca
     2101 tgaaaggaat atctgcttca tattttttat cattacaagg catgatagta actgtataaa
     2161 catcctcgag ctgcagtata aaatataaat aattttctaa aaaacttaac ttcatgtgaa
     2221 aagtttgtta aaatataaat gagcacgtta atcatttaac atagataatt aaatagtaaa
     2281 agggagtgtc gacccgggat ggataagaaa tactcaatag gcttagctat cggcacaaat
     2341 agcgtcggat gggcggtgat cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt
     2401 ctgggaaata cagaccgcca cagtatcaaa aaaaatctta taggggctct tttatttgac
     2461 agtggagaga cagcggaagc gactcgtctc aaacggacag ctcgtagaag gtatacacgt
     2521 cggaagaatc gtatttgtta tctacaggag attttttcaa atgagatggc gaaagtagat
     2581 gatagtttct ttcatcgact tgaagagtct tttttggtgg aagaagacaa gaagcatgaa
     2641 cgtcatccta tttttggaaa tatagtagat gaagttgctt atcatgagaa atatccaact
     2701 atctatcatc tgcgaaaaaa attggtagat tctactgata aagcggattt gcgcttaatc
     2761 tatttggcct tagcgcatat gattaagttt cgtggtcatt ttttgattga gggagattta
     2821 aatcctgata atagtgatgt ggacaaacta tttatccagt tggtacaaac ctacaatcaa
     2881 ttatttgaag aaaaccctat taacgcaagt ggagtagatg ctaaagcgat tctttctgca
     2941 cgattgagta aatcaagacg attagaaaat ctcattgctc agctccccgg tgagaagaaa
     3001 aatggcttat ttgggaatct cattgctttg tcattgggtt tgacccctaa ttttaaatca
     3061 aattttgatt tggcagaaga tgctaaatta cagctttcaa aagatactta cgatgatgat
     3121 ttagataatt tattggcgca aattggagat caatatgctg atttgttttt ggcagctaag
     3181 aatttatcag atgctatttt actttcagat atcctaagag taaatactga aataactaag
     3241 gctcccctat cagcttcaat gattaaacgc tacgatgaac atcatcaaga cttgactctt
     3301 ttaaaagctt tagttcgaca acaacttcca gaaaagtata aagaaatctt ttttgatcaa
     3361 tcaaaaaacg gatatgcagg ttatattgat gggggagcta gccaagaaga attttataaa
     3421 tttatcaaac caattttaga aaaaatggat ggtactgagg aattattggt gaaactaaat
     3481 cgtgaagatt tgctgcgcaa gcaacggacc tttgacaacg gctctattcc ccatcaaatt
     3541 cacttgggtg agctgcatgc tattttgaga agacaagaag acttttatcc atttttaaaa
     3601 gacaatcgtg agaagattga aaaaatcttg acttttcgaa ttccttatta tgttggtcca
     3661 ttggcgcgtg gcaatagtcg ttttgcatgg atgactcgga agtctgaaga aacaattacc
     3721 ccatggaatt ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc
     3781 atgacaaact ttgataaaaa tcttccaaat gaaaaagtac taccaaaaca tagtttgctt
     3841 tatgagtatt ttacggttta taacgaattg acaaaggtca aatatgttac tgaaggaatg
     3901 cgaaaaccag catttctttc aggtgaacag aagaaagcca ttgttgattt actcttcaaa
     3961 acaaatcgaa aagtaaccgt taagcaatta aaagaagatt atttcaaaaa aatagaatgt
     4021 tttgatagtg ttgaaatttc aggagttgaa gatagattta atgcttcatt aggtacctac
     4081 catgatttgc taaaaattat taaagataaa gattttttgg ataatgaaga aaatgaagat
     4141 atcttagagg atattgtttt aacattgacc ttatttgaag atagggagat gattgaggaa
     4201 agacttaaaa catatgctca cctctttgat gataaggtga tgaaacagct taaacgtcgc
     4261 cgttatactg gttggggacg tttgtctcga aaattgatta atggtattag ggataagcaa
     4321 tctggcaaaa caatattaga ttttttgaaa tcagatggtt ttgccaatcg caattttatg
     4381 cagctgatcc atgatgatag tttgacattt aaagaagaca ttcaaaaagc acaagtgtct
     4441 ggacaaggcg atagtttaca tgaacatatt gcaaatttag ctggtagccc tgctattaaa
     4501 aaaggtattt tacagactgt aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat
     4561 aagccagaaa atatcgttat tgaaatggca cgtgaaaatc agacaactca aaagggccag
     4621 aaaaattcgc gagagcgtat gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag
     4681 attcttaaag agcatcctgt tgaaaatact caattgcaaa atgaaaagct ctatctctat
     4741 tatctccaaa atggaagaga catgtatgtg gaccaagaat tagatattaa tcgtttaagt
     4801 gattatgatg tcgatcacat tgttccacaa agtttcctta aagacgattc aatagacaat
     4861 aaggtcttaa cgcgttctga taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa
     4921 gtagtcaaaa agatgaaaaa ctattggaga caacttctaa acgccaagtt aatcactcaa
     4981 cgtaagtttg ataatttaac gaaagctgaa cgtggaggtt tgagtgaact tgataaagct
     5041 ggttttatca aacgccaatt ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt
     5101 ttggatagtc gcatgaatac taaatacgat gaaaatgata aacttattcg agaggttaaa
     5161 gtgattacct taaaatctaa attagtttct gacttccgaa aagatttcca attctataaa
     5221 gtacgtgaga ttaacaatta ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga
     5281 actgctttga ttaagaaata tccaaaactt gaatcggagt ttgtctatgg tgattataaa
     5341 gtttatgatg ttcgtaaaat gattgctaag tctgagcaag aaataggcaa agcaaccgca
     5401 aaatatttct tttactctaa tatcatgaac ttcttcaaaa cagaaattac acttgcaaat
     5461 ggagagattc gcaaacgccc tctaatcgaa actaatgggg aaactggaga aattgtctgg
     5521 gataaagggc gagattttgc cacagtgcgc aaagtattgt ccatgcccca agtcaatatt
     5581 gtcaagaaaa cagaagtaca gacaggcgga ttctccaagg agtcaatttt accaaaaaga
     5641 aattcggaca agcttattgc tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt
     5701 gatagtccaa cggtagctta ttcagtccta gtggttgcta aggtggaaaa agggaaatcg
     5761 aagaagttaa aatccgttaa agagttacta gggatcacaa ttatggaaag aagttccttt
     5821 gaaaaaaatc cgattgactt tttagaagct aaaggatata aggaagttaa aaaagactta
     5881 atcattaaac tacctaaata tagtcttttt gagttagaaa acggtcgtaa acggatgctg
     5941 gctagtgccg gagaattaca aaaaggaaat gagctggctc tgccaagcaa atatgtgaat
     6001 tttttatatt tagctagtca ttatgaaaag ttgaagggta gtccagaaga taacgaacaa
     6061 aaacaattgt ttgtggagca gcataagcat tatttagatg agattattga gcaaatcagt
     6121 gaattttcta agcgtgttat tttagcagat gccaatttag ataaagttct tagtgcatat
     6181 aacaaacata gagacaaacc aatacgtgaa caagcagaaa atattattca tttatttacg
     6241 ttgacgaatc ttggagctcc cgctgctttt aaatattttg atacaacaat tgatcgtaaa
     6301 cgatatacgt ctacaaaaga agttttagat gccactctta tccatcaatc catcactggt
     6361 ctttatgaaa cacgcattga tttgagtcag ctaggaggtg actgacccgg gaattcactg
     6421 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     6481 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     6541 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     6601 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     6661 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6721 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     6781 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     6841 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     6901 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6961 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7021 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7081 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7141 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7201 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     7261 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7321 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7381 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7441 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7501 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7561 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7621 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7681 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7741 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7801 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7861 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7921 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7981 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8041 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8101 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8161 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     8221 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8281 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8341 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8401 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8461 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8521 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8581 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8641 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8701 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8761 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     8821 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     8881 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8941 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     9001 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ttggctaaca
     9061 cacacgccat tccaaccaat agttttctcg gcataaagcc atgctctgac gcttaaatgc
     9121 actaatgcct taaaaaaaca ttaaagtcta acacactaga cttatttact tcgtaattaa
     9181 gtcgttaaac cgtgtgctct acgaccaaaa gtataaaacc tttaagaact ttcttttttc
     9241 ttgtaaaaaa agaaactaga taaatctctc atatctttta ttcaataatc gcatcagatt
     9301 gcagtataaa tttaacgatc actcatcatg ttcatattta tcagagctcg tgctataatt
     9361 atactaattt tataaggagg aaaaaataaa gagggttata atgaacgaga aaaatataaa
     9421 acacagtcaa aactttatta cttcaaaaca taatatagat aaaataatga caaatataag
     9481 attaaatgaa catgataata tctttgaaat cggctcagga aaagggcatt ttacccttga
     9541 attagtacag aggtgtaatt tcgtaactgc cattgaaata gaccataaat tatgcaaaac
     9601 tacagaaaat aaacttgttg atcacgataa tttccaagtt ttaaacaagg atatattgca
     9661 gtttaaattt cctaaaaacc aatcctataa aatatttggt aatatacctt ataacataag
     9721 tacggatata atacgcaaaa ttgtttttga tagtatagct gatgagattt atttaatcgt
     9781 ggaatacggg tttgctaaaa gattattaaa tacaaaacgc tcattggcat tatttttaat
     9841 ggcagaagtt gatatttcta tattaagtat ggttccaaga gaatattttc atcctaaacc
     9901 taaagtgaat agctcactta tcagattaaa tagaaaaaaa tcaagaatat cacacaaaga
     9961 taaacagaag tataattatt tcgttatgaa atgggttaac aaagaataca agaaaatatt
    10021 tacaaaaaat caatttaaca attccttaaa acatgcagga attgacgatt taaacaatat
    10081 tagctttgaa caattcttat ctcttttcaa tagctataaa ttatttaata agtaagttaa
    10141 gggatgcata aactgcatcc cttaacttgt ttttcgtgta cctatttttt gtgaatcgat
    10201 tatgtctttt gcgcattcac ttcttttcta tataaatatg agcgaagcga ataagcgtcg
    10261 gaaaagcagc aaaaagtttc ctttttgctg ttggagcatg ggggttcagg gggtgcagta
    10321 tctgacgtca atgccgagcg aaagcgagcc gaagggtagc atttacgtta gataaccccc
    10381 tgatatgctc cgacgcttta tatagaaaag aagattcaac taggtaaaat cttaatatag
    10441 gttgagatga taaggtttat aaggaatttg tttgttctaa tttttcactc attttgttct
    10501 aatttctttt aacaaatgtt cttttttttt tagaacagtt atgatatagt tagaatagtt
    10561 taaaataagg agtgagaaaa agatgaaaga aagatatgga acagtctata aaggctctca
    10621 gaggctcata gacgaagaaa gtggagaagt catagaggta gacaagttat accgtaaaca
    10681 aacgtctggt aacttcgtaa aggcatatat agtgcaatta ataagtatgt tagatatgat
    10741 tggcggaaaa aaacttaaaa tcgttaacta tatcctagat aatgtccact taagtaacaa
    10801 tacaatgata gctacaacaa gagaaatagc aaaagctaca ggaacaagtc tacaaacagt
    10861 aataacaaca cttaaaatct tagaagaagg aaatattata aaaagaaaaa ctggagtatt
    10921 aatgttaaac cctgaactac taatgagagg cgacgaccaa aaacaaaaat acctcttact
    10981 cgaatttggg aactttgagc aagaggcaaa tgaaatagat tgacctccca ataacaccac
    11041 gtagttattg ggaggtcaat ctatgaaaat gcgatta
//
