LOCUS       Exported                3444 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 3444)
  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..3444
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=JI601_hU6_SaCas9gRNA(updated)_RIP"
     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..1498
                     /note="Rat Ins1 promoter (RIP)"
     misc_recomb     1506..1663
                     /label=attL5r
                     /note="attL5r recombination site"
                     /note="recognized as attR5"
     misc_feature    join(1514..3444,1..658)
                     /label=nonstandard type: vector
                     /note="/source=pDONR221-P1P5r"
                     /note="/type=Donor Vector"
     primer_bind     complement(1709..1725)
                     /label=M13 Reverse primer binding site
                     /note="M13 Reverse primer binding site"
     gene            1838..2647
                     /label=Kanamycin resistance gene
                     /note="Kanamycin resistance gene"
     rep_origin      2768..3441
                     /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 agaaagactg agctaagaat
     1081 ccagctatca atagaaacta tgaaacagtt ccagggacaa agataccagg tccccaacaa
     1141 ctgcaacttt ctgggaaatg aggtggaaaa tgctcagcca aggaaaaaga gggccttacc
     1201 ctctctggga caatgattgt gctgtgaact gcttcatcag gccatctggc cccttgttaa
     1261 taatctaatt accctaggtc taagtagagt tgttgacgtc caatgagcgc tttctgcaga
     1321 cttagcacta ggcaagtgtt tggaaattac agcttcagcc cctctcgcca tctgcctacc
     1381 tacccctcct agagccctta atgggccaaa cggcaaagtc cagggggcag agaggaggtg
     1441 ctttggacta taaagctagt ggggacccag taactcccaa ccctaagtga ccagctacga
     1501 attccacaac tttgtataca aaagttgaac gagaaacgta aaatgatata aatatcaata
     1561 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     1621 agtcactatg aatcaactac ttagatggta ttagtgacct gtactgcagg gcggccgcga
     1681 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1741 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1801 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1861 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1921 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1981 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2041 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2101 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2161 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2221 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2281 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2341 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2401 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2461 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2521 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2581 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2641 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2701 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3421 ttgctggcct tttgctcaca tgtt
//
