LOCUS       Exported                3772 bp ds-DNA     circular SYN 17-JUL-2006
DEFINITION  Complementary copy of pDONR221-P1P5r.
ACCESSION   .
VERSION     .
KEYWORDS    pDONR221-P1P5r
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3772)
  AUTHORS   Invitrogen
  TITLE     Direct Submission
  JOURNAL   Exported Jan 17, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3772
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=KB501_hU6_SaCas9gRNA(updated)_CMV"
     misc_feature    complement(268..295)
                     /label=rrnB T2 transcription termination sequence
                     /note="rrnB T2 transcription termination sequence"
     misc_feature    complement(427..470)
                     /label=rrnB T1 transcription termination sequence
                     /note="rrnB T1 transcription termination sequence"
     primer_bind     517..533
                     /label=M13 Forward (-40) primer binding site
                     /note="M13 Forward (-40) primer binding site"
     primer_bind     537..552
                     /label=M13 Forward (-20) primer binding site
                     /note="M13 Forward (-20) primer binding site"
     misc_recomb     570..668
                     /label=attL1
                     /note="attL1 recombination site"
                     /note="recognized as attL1"
     promoter        679..927
                     /note="human U6 promoter"
     misc_feature    924..947
                     /label=BsaI sense fragment
                     /note="BsaI sense fragment"
                     /note="Replace with sense protospacer motif with sequence 
                     5%92-CACC(G)N(20-22)-3%92"
     misc_feature    complement(928..951)
                     /label=BsaI antisense fragment
                     /note="BsaI antisense fragment"
                     /note="Replace with antisense protospacer motif with 
                     sequence 5%92-AAACN(20-22)(C)-3%92"
     misc_RNA        948..1037
                     /label=SaCas9 sgRNA scaffold
                     /note="SaCas9 sgRNA scaffold"
                     /note="adopted from PMID: 26721686 and PMID: 27210042"
     promoter        1064..1822
                     /note="CMV promoter"
     misc_recomb     1834..1991
                     /label=attL5r
                     /note="attL5r recombination site"
                     /note="recognized as attR5"
     misc_feature    join(1842..3772,1..658)
                     /label=nonstandard type: vector
                     /note="/source=pDONR221-P1P5r"
                     /note="/type=Donor Vector"
     primer_bind     complement(2037..2053)
                     /label=M13 Reverse primer binding site
                     /note="M13 Reverse primer binding site"
     gene            2166..2975
                     /label=Kanamycin resistance gene
                     /note="Kanamycin resistance gene"
     rep_origin      3096..3769
                     /label=f1 origin
                     /note="f1 origin"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctta tggtacctga gggcctattt cccatgattc cttcatattt gcatatacga
      721 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
      781 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
      841 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
      901 tatatcttgt ggaaaggacg aaacaccgga gaccacggca ggtctcagtt taagtactct
      961 gtgctggaaa cagcacagaa tctactaaac aaggcaaaat gccgtgttta tctcgtcaac
     1021 ttgttggcga gatttttgcg gccgctgtac atacgactct agaacgatct atacattgaa
     1081 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     1141 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     1201 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     1261 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     1321 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1381 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1441 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
     1501 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
     1561 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
     1621 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     1681 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
     1741 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1801 tcgtttagtg aaccgtcaga tccggcgaat tccacaactt tgtatacaaa agttgaacga
     1861 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     1921 acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt agatggtatt
     1981 agtgacctgt actgcagggc ggccgcgata tcccctatag tgagtcgtat tacatggtca
     2041 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2101 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2161 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2221 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2281 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2341 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2401 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2461 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2521 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2581 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2641 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2701 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2761 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2821 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2881 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2941 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     3001 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     3061 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3121 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3181 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3241 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3601 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3661 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3721 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
