LOCUS       pDONR221-P1P5r          4765 bp    DNA     circular SYN 23-APR-2009
DEFINITION  Complementary copy of pDONR221-P1P5r.
SOURCE      
  ORGANISM  
COMMENT     VNTDATE|399123546|
COMMENT     VNTDBDATE|416243884|
COMMENT     VNTNAME|pDONR221-P1P5r|
COMMENT     VNTAUTHORNAME|Invitrogen|
COMMENT     VNTAUTHORTEL|(800)-357-3114|
COMMENT     VNTAUTHORFAX|(760)-602-6500|
COMMENT     VNTOAUTHORFTP|ftp://ftp.bioinformatics.invitrogen.com|
COMMENT     VNTAUTHOREML|bioinfosales@invitrogen.com|
COMMENT     VNTAUTHORWWW|http://www.invitrogen.com/bioinformatics|
COMMENT     VNTAUTHORAD1|Invitrogen Corporation|
COMMENT     VNTAUTHORAD2|1620 Faraday Avenue|
COMMENT     VNTAUTHORAD3|Carlsbad, CA 92008 U.S.A.|
FEATURES             Location/Qualifiers
     misc_feature    complement(427..470)
                     /note="rrnB T1 transcription termination sequence"
     misc_feature    complement(268..295)
                     /note="rrnB T2 transcription termination sequence"
     primer_bind     537..552
                     /note="M13 Forward (-20) primer binding site"
     primer_bind     517..533
                     /note="M13 Forward (-40) primer binding site"
     primer_bind     complement(1317..1333)
                     /note="M13 Reverse primer binding site"
     rep_origin      2376..3049
                     /note="f1 origin"
     gene            1446..2255
                     /note="Kanamycin resistance gene"
     misc_recomb     570..668
                     /label=attL1
                     /note="attL1 recombination site"
                     /note="recognized as attL1"
     misc_recomb     1114..1271
                     /label=attL5r
                     /note="attL5r recombination site"
                     /note="recognized as attR5"
     promoter        670..984
                     /note="mouse U6 promoter"
     vector          join(1122..3052,1..658)
                     /source="pDONR221-P1P5r"
                     /type="Donor Vector"
     misc_RNA        1005..1096
                     /note="SpCas9 optimized gRNA"
                     /note="also compatible w/ x-Cas9"
                     /note="Based on PMID: 26671237 and PMID: 27210042"
     misc_feature    981..1004
                     /note="BsaI sense fragment"
                     /note="Replace with sense protospacer motif with sequence
                     5%92-TGTTT(G)N(19)-3%92"
     misc_feature    complement(985..1008)
                     /note="BsaI antisense fragment"
                     /note="Replace with antisense protospacer motif with
                     sequence 5%92-AAACN(19)(C)A-3%92"
     source          1..3052
                     /dnas_title="LG001_msU6_SpCas9_gRNA"
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 GATCCGACGC CGCCATCTCT AGGCCCGCGC CGGCCCCCTC GCACAGACTT
      721 GTGGGAGAAG CTCGGCTACT CCCCTGCCCC GGTTAATTTG CATATAATAT TTCCTAGTAA
      781 CTATAGAGGC TTAATGTGCG ATAAAAGACA GATAATCTGT TCTTTTTAAT ACTAGCTACA
      841 TTTTACATGA TAGGCTTGGA TTTCTATAAG AGATACAAAT ACTAAATTAT TATTTTAAAA
      901 AACAGCACAA AAGGAAACTC ACCCTAACTG TAAAGTAATT GTGTGTTTTG AGACTATAAA
      961 TATCCCTTGG AGAAAAGCCT TGTTgGAGAC CacggcaGGT CTCagtttca gagctatgct
     1021 ggaaacagca tagcaagttg aaataaggct agtccgttat caacttgaaa aagtggcacc
     1081 gagtcggtgc ttttttgcgg ccgcaggaag cttACAACTT Tgtatacaaa agttgaacga
     1141 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     1201 acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt agatggtatt
     1261 agtgacctgt actgcagggc ggccgcgata tcccctatag tgagtcgtat tacatggtca
     1321 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     1381 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1441 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     1501 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1561 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1621 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1681 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1741 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1801 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1861 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1921 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1981 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2041 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2101 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2161 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2221 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     2281 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     2341 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2401 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2461 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2521 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2581 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2641 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2701 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2761 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2821 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2881 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2941 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3001 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//

