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(1314..1330)
                     /note="M13 Reverse primer binding site"
     rep_origin      2373..3046
                     /note="f1 origin"
     gene            1443..2252
                     /note="Kanamycin resistance gene"
     misc_recomb     570..668
                     /label=attL1
                     /note="attL1 recombination site"
                     /note="recognized as attL1"
     misc_recomb     1111..1268
                     /label=attL5r
                     /note="attL5r recombination site"
                     /note="recognized as attR5"
     promoter        670..984
                     /note="mouse U6 promoter"
     vector          join(1119..3049,1..658)
                     /source="pDONR221-P1P5r"
                     /type="Donor Vector"
     misc_RNA        1005..1094
                     /note="SaCas9 sgRNA scaffold"
                     /note="adopted from  PMID: 26721686 and PMID: 27210042 "
     misc_feature    981..1004
                     /note="BsaI sense fragment"
                     /note="Replace with sense protospacer motif with sequence
                     5%92-TGTTT(G)N(20-22)-3%92"
     misc_feature    complement(985..1008)
                     /note="BsaI antisense fragment"
                     /note="Replace with antisense protospacer motif with
                     sequence 5%92-AAACN(20-22)(C)A-3%92"
     source          1..3049
                     /dnas_title="LF901_msU6_SaCas9_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 CTCagtttAa gtactctgTG
     1021 CTGgaaaCAG CAcagaatct actaaacaag gcaaaatgcc gtgtttatct cgtcaacttg
     1081 ttggcgagat ttttgcggcc gctgaagctt ACAACTTTgt atacaaaagt tgaacgagaa
     1141 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     1201 taatactgta aaacacaaca tatccagtca ctatgaatca actacttaga tggtattagt
     1261 gacctgtact gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     1321 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     1381 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1441 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     1501 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     1561 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     1621 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     1681 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     1741 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1801 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1861 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1921 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1981 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2041 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2101 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2161 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2221 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     2281 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     2341 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2401 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2521 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2581 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2641 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3001 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//

