LOCUS       Exported               11707 bp ds-DNA     circular SYN 28-Ò»ÔÂ-2016
DEFINITION  synthetic circular DNA
ACCESSION   pIMP1
VERSION     .
KEYWORDS    pNICKclos2.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11707)
  AUTHORS   Self
  JOURNAL   Unpublished.
REFERENCE   2  (bases 1 to 11707)
  AUTHORS   SZ
  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..11707
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..1485
                     /note="pCB102"
     misc_feature    1487..1623
                     /note="sgRNA"
     misc_feature    1487..1521
                     /note="Pj23119"
     misc_feature    1522..1541
                     /note="20bp spacer"
     misc_feature    1542..1623
                     /note="tracrRNA"
     misc_feature    1632..2631
                     /note="cbei2385-HA-up"
     misc_feature    2638..3637
                     /note="cbei2385-HA-dn"
     misc_feature    3644..3796
                     /note="Pthl"
     misc_feature    3728..3746
                     /note="seq-s"
     misc_feature    3808..7914
                     /note="cas9-nickase"
     misc_feature    7984..8002
                     /note="seq-a"
     CDS             8523..9380
                     /codon_start=1
                     /gene="bla"
                     /product="B-lactamase"
                     /note="bla"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    10141..10203
                     /note="ori"
     CDS             10912..11643
                     /codon_start=1
                     /gene="mslR"
                     /note="mslR"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVQRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIFGNIPY
                     NISTDIIRKIVFDSIADEIYLIVEYGFAKRLLNTKRSLALFLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLNRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
ORIGIN
        1 cgataattta cagaaaagaa aattatagaa tttagtatga ttaattatac tcatttatga
       61 atgtttaatt gaatacaaaa aaaaatactt gttatgtatt caattacggg ttaaaatata
      121 gacaagttga aaaatttaat aaaaaaataa gtcctcagct cttatatatt aagctaccaa
      181 cttagtatat aagccaaaac ttaaatgtgc taccaacaca tcaagccgtt agagaactct
      241 atctatagca atatttcaaa tgtaccgaca tacaagagaa acattaacta tatatattca
      301 atttatgaga ttatcttaac agatataaat gtaaattgca ataagtaaga tttagaagtt
      361 tatagccttt gtgtattgga agcagtacgc aaaggctttt ttatttgata aaaattagaa
      421 gtatatttat tttttcataa ttaatttatg aaaatgaaag ggggtgagca aagtgacaga
      481 ggaaagcagt atcttatcaa ataacaaggt attagcaata tcattattga ctttagcagt
      541 aaacattatg acttttatag tgcttgtagc taagtagtac gaaaggggga gctttaaaaa
      601 gctccttgga atacatagaa ttcataaatt aatttatgaa aagaagggcg tatatgaaaa
      661 cttgtaaaaa ttgcaaagag tttattaaag atactgaaat atgcaaaata cattcgttga
      721 tgattcatga taaaacagta gcaacctatt gcagtaaata caatgagtca agatgtttac
      781 ataaagggaa agtccaatgt attaattgtt caaagatgaa ccgatatgga tggtgtgcca
      841 taaaaatgag atgttttaca gaggaagaac agaaaaaaga acgtacatgc attaaatatt
      901 atgcaaggag ctttaaaaaa gctcatgtaa agaagagtaa aaagaaaaaa taatttattt
      961 attaatttaa tattgagagt gccgacacag tatgcactaa aaaatatatc tgtggtgtag
     1021 tgagccgata caaaaggata gtcactcgca ttttcataat acatcttatg ttatgattat
     1081 gtgtcggtgg gacttcacga cgaaaaccca caataaaaaa agagttcggg gtagggttaa
     1141 gcatagttga ggcaactaaa caatcaagct aggatatgca gtagcagacc gtaaggtcgt
     1201 tgtttaggtg tgttgtaata catacgctat taagatgtaa aaatacggat accaatgaag
     1261 ggaaaagtat aatttttgga tgtagtttgt ttgttcatct atgggcaaac tacgtccaaa
     1321 gccgtttcca aatctgctaa aaagtatatc ctttctaaaa tcaaagtcaa gtatgaaatc
     1381 ataaataaag tttaattttg aagttattat gatattatgt ttttctatta aaataaatta
     1441 agtatataga atagtttaat aatagtatat acttaatgtg ctgcagttga cagctagctc
     1501 agtcctaggt ataatactag tcgagttaga cataatagtg agttttagag ctagaaatag
     1561 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     1621 tttgcggccg caaaacctgg ttctaatgga attgtttact taccatatct tatgggagaa
     1681 agaacacctc atatagaccc taatgtaaaa ggtgcatttt taggagtatc actaataaat
     1741 aaccatgatg attttgtacg tagtatatta gaaggtgtta gctttagttt gaaaaactgc
     1801 cttgatttaa tcgaagacat gaatgtaaac attgatgaaa taagagtaag tggcggtgga
     1861 gcagaaagtg ctgtatggag acaaatatta gctgatattt tccaatattc attaactaca
     1921 gttaaagcat ctgaaggcgg tgcgcttgga gttgcaatat tagcaggggt tggagcagga
     1981 atctatgatt ctgtagaaga tgcttgtgat agaatcgtaa aaggaaacaa gaaagtttct
     2041 ccaaatgaag aattaaaaga agtatactca aaagtatatg aaacatataa ttcagcttat
     2101 ccaagaataa aagatattta aatatattaa agatagaaat caatcatctt agttaagtaa
     2161 ttgtataatt aattaagggt gtgtagtagt atcgaatttt agcaaaaaaa tcatatagtt
     2221 ttttaaatac tttaacaaaa ataataatag tgatattatt taagaaaaag ctactattac
     2281 tgataatagt cgagaggagt attttaaagt ttgatagaag taaatcacag taaaataaaa
     2341 gaaactaaca gaaaaaagat aatagcatta ttattaaaga aaaatgagat caccaaatta
     2401 gatatatcta gaatattaga tataagtata acaacagttt ctacaaatat tacagaattg
     2461 aaaagcgaag gtattgtaga agatgtaaga tcgttggaat ctacaggagg aagaaaagca
     2521 attgctatta agttgaatga aaactgtaga tattcaattg gagtcgcact aacaccgaat
     2581 catataaaga tatcattagt aaatataaag aaaaaagtta ttgaaagtat gctgcagtgg
     2641 aatagagaac ataataaatt tacttaatga aaatatagat ttgttaatga agaaatataa
     2701 tttatgctct gcaaatttat taggaatagg gatttcatta cctggaaccg tagattttaa
     2761 agaaggcata ataaagtatt catatttgtt aggcgccaaa gattttaatt taaaggaaaa
     2821 gttcgaatac ttagacattc cagtatatgt agataatgaa gcaaatttat ctgcttacta
     2881 cgagttttta aataaaagag atatactaag aaatctatta tatgtatcaa taacagaagg
     2941 tttaggcctt ggtataataa taaatggaaa aatttacaga ggtgacaata actcatcagg
     3001 agaattaggt catactaaga tagcaattga tggaaaaaaa tgtaagtgtg gagctagagg
     3061 ctgtttagaa gcatacacat ctatgaattc tctgattgat agttataatg aagctaatag
     3121 ttcaaatatt agtgatattg atgaatttga agaaagatat aatcaaaatg acaaagatgc
     3181 tcatgatgta ttaaatgatt atttaaaaac attgtcttta ggtctatcga atttggttat
     3241 gatatttgat cctagcagca tagtaatagg tggagatata aataatttac ttaatgataa
     3301 gatagatatg ctgaaaaaag aggtttataa agataattta tttacaaatg aaaataattg
     3361 cagtataagt attgcaagtt ttaaagaatc ttatctacta ggagctgcta tgatgcctat
     3421 agaagaattc ttagaaatta aataatggag tggtgaaagc tgctctatta atttaaaaac
     3481 ttattaaata aatttttaaa atatattttt aaagttatgt aagcatagct atattaaggg
     3541 gggtcttaat ctttgattgg aactccagat taagacaagc cttttatata aaactaaata
     3601 tctatttatt tcatattaga tttatgttct tgattttctc gagtttttaa caaaatatat
     3661 tgataaaaat aataatagtg ggtataatta agttgttaga gaaaacgtat aaattaggga
     3721 taaactatgg aacttatgaa atagattgaa atggtttatc tgttaccccg tatcaaaatt
     3781 taggaggtta gttagaggat ccccgggatg gataagaaat actcaatagg cttagctatc
     3841 ggcacaaata gcgtcggatg ggcggtgatc actgatgaat ataaggttcc gtctaaaaag
     3901 ttcaaggttc tgggaaatac agaccgccac agtatcaaaa aaaatcttat aggggctctt
     3961 ttatttgaca gtggagagac agcggaagcg actcgtctca aacggacagc tcgtagaagg
     4021 tatacacgtc ggaagaatcg tatttgttat ctacaggaga ttttttcaaa tgagatggcg
     4081 aaagtagatg atagtttctt tcatcgactt gaagagtctt ttttggtgga agaagacaag
     4141 aagcatgaac gtcatcctat ttttggaaat atagtagatg aagttgctta tcatgagaaa
     4201 tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa agcggatttg
     4261 cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt tttgattgag
     4321 ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt ggtacaaacc
     4381 tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc taaagcgatt
     4441 ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca gctccccggt
     4501 gagaagaaaa atggcttatt tgggaatctc attgctttgt cattgggttt gacccctaat
     4561 tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa agatacttac
     4621 gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga tttgtttttg
     4681 gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt aaatactgaa
     4741 ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca tcatcaagac
     4801 ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa agaaatcttt
     4861 tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag ccaagaagaa
     4921 ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga attattggtg
     4981 aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg ctctattccc
     5041 catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga cttttatcca
     5101 tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat tccttattat
     5161 gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa gtctgaagaa
     5221 acaattaccc catggaattt tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt
     5281 attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact accaaaacat
     5341 agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa atatgttact
     5401 gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat tgttgattta
     5461 ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa
     5521 atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa tgcttcatta
     5581 ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga taatgaagaa
     5641 aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga tagggagatg
     5701 attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat gaaacagctt
     5761 aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa tggtattagg
     5821 gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt tgccaatcgc
     5881 aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat tcaaaaagca
     5941 caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc tggtagccct
     6001 gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt caaagtaatg
     6061 gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca gacaactcaa
     6121 aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat caaagaatta
     6181 ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc
     6241 tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt agatattaat
     6301 cgtttaagtg attatgatgt cgatcacatt gttccacaaa gtttccttaa agacgattca
     6361 atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca
     6421 agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa cgccaagtta
     6481 atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt gagtgaactt
     6541 gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac taagcatgtg
     6601 gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa acttattcga
     6661 gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa agatttccaa
     6721 ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta tctaaatgcc
     6781 gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt tgtctatggt
     6841 gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa
     6901 gcaaccgcaa aatatttctt ttactctaat atcatgaact tcttcaaaac agaaattaca
     6961 cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga aactggagaa
     7021 attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc catgccccaa
     7081 gtcaatattg tcaagaaaac agaagtacag acaggcggat tctccaagga gtcaatttta
     7141 ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc aaaaaaatat
     7201 ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa
     7261 gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat tatggaaaga
     7321 agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa ggaagttaaa
     7381 aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa cggtcgtaaa
     7441 cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct gccaagcaaa
     7501 tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag tccagaagat
     7561 aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga gattattgag
     7621 caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga taaagttctt
     7681 agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa tattattcat
     7741 ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga tacaacaatt
     7801 gatcgtaaac gatatacgtc tacaaaagaa gttttagatg ccactcttat ccatcaatcc
     7861 atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga ctgagcggcc
     7921 gcaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     7981 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     8041 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     8101 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     8161 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     8221 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     8281 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     8341 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     8401 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     8461 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     8521 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     8581 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     8641 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     8701 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     8761 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     8821 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     8881 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     8941 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     9001 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     9061 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     9121 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     9181 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     9241 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     9301 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     9361 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     9421 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     9481 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     9541 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     9601 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     9661 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     9721 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     9781 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     9841 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     9901 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     9961 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    10021 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    10081 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    10141 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    10201 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    10261 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    10321 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    10381 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    10441 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    10501 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    10561 tttggctaac acacacgcca ttccaaccaa tagttttctc ggcataaagc catgctctga
    10621 cgcttaaatg cactaatgcc ttaaaaaaac attaaagtct aacacactag acttatttac
    10681 ttcgtaatta agtcgttaaa ccgtgtgctc tacgaccaaa agtataaaac ctttaagaac
    10741 tttctttttt cttgtaaaaa aagaaactag ataaatctct catatctttt attcaataat
    10801 cgcatcagat tgcagtataa atttaacgat cactcatcat gttcatattt atcagagctc
    10861 gtgctataat tatactaatt ttataaggag gaaaaaataa agagggttat aatgaacgag
    10921 aaaaatataa aacacagtca aaactttatt acttcaaaac ataatataga taaaataatg
    10981 acaaatataa gattaaatga acatgataat atctttgaaa tcggctcagg aaaagggcat
    11041 tttacccttg aattagtaca gaggtgtaat ttcgtaactg ccattgaaat agaccataaa
    11101 ttatgcaaaa ctacagaaaa taaacttgtt gatcacgata atttccaagt tttaaacaag
    11161 gatatattgc agtttaaatt tcctaaaaac caatcctata aaatatttgg taatatacct
    11221 tataacataa gtacggatat aatacgcaaa attgtttttg atagtatagc tgatgagatt
    11281 tatttaatcg tggaatacgg gtttgctaaa agattattaa atacaaaacg ctcattggca
    11341 ttatttttaa tggcagaagt tgatatttct atattaagta tggttccaag agaatatttt
    11401 catcctaaac ctaaagtgaa tagctcactt atcagattaa atagaaaaaa atcaagaata
    11461 tcacacaaag ataaacagaa gtataattat ttcgttatga aatgggttaa caaagaatac
    11521 aagaaaatat ttacaaaaaa tcaatttaac aattccttaa aacatgcagg aattgacgat
    11581 ttaaacaata ttagctttga acaattctta tctcttttca atagctataa attatttaat
    11641 aagtaagtta agggatgcat aaactgcatc ccttaacttg tttttcgtgt acctattttt
    11701 tgtgaat
//
