LOCUS       pENTR207-PRS-mRIP1-D156A(KD+RHIM)      5062 bp    DNA     circular UNA 10-FEB-2012
DEFINITION  LMBP 8818.
SOURCE
  ORGANISM
COMMENT     Accession number: LMBP 8818
            (c) BCCM/GeneCorner Plasmid Collection (Ghent University, Belgium)
FEATURES             Location/Qualifiers
     CDS             1..1665
                     /label="mRIP1D156A ( KD + RHIM ) + stop"
     Misc._feature   complement(1670..1765)
                     /label=attL2
     Unsure          1677
                     /Design_Description="If necessary, linearize attB sequence
                     mRIP1D156A  fragment met primers voor constructie 204 at a
                     unique REN site outside the attB region. Ethanol
                     precipitate and resuspend.	Mix linearized attB sequence
                     mRIP1D156A  fragment met primers voor constructie 204 and
                     pDONR Vector pDONR207 in BP buffer and TE buffers.	Thaw,
                     vortex and add BP Clonase enzyme. Incubate >= 1 hr. at
                     25°.	Add Proteinase K for 10 min at 37°.	Transform into
                     DH5a or TOP10 E. coli or any non-F' episome bearing
                     strain.	Select Entry Clones on agar plates with antibiotic
                     gentamicin."
                     /label="Gateway Cloning Description"
     Insertion       1678..5020
                     /note="Region on donor sequence (pDONR207): 2657..414"
                     /label=pDONR207
     Primer          complement(1790..1813)
                     /invitrogen="1994000"
                     /label="pDONR?E07 reverse primer"
     CDS             complement(2549..3082)
                     /invitrogen="1150000"
                     /label=Gm(R)
     Promoter        complement(3271..3298)
                     /invitrogen="2250000"
                     /label="Pc promoter"
     Terminator      4679..4706
                     /invitrogen="2590000"
                     /label="rrnB T2 transcription terminator"
     Terminator      4838..4881
                     /invitrogen="2580000"
                     /label="rrnB T1 transcription terminator"
     Primer          4906..4930
                     /invitrogen="1993000"
                     /label="pDONR?E07 forward primer"
     Misc._feature   4938..5033
                     /label=attL1
     Insertion       5021..>5062
                     /note="Region on donor sequence (mRIP1D156A  fragment met
                     primers voor constructie 204): 14..1732"
                     /label="mRIP1D156A  fragment met primers voor constructie 204"
     Cleavage_site   5039..5062
                     /label="prescission cleavage site ( PRS)"
ORIGIN
        1 ATGCAACCAG ACATGTCCTT GGACAATATT AAGATGGCAT CCAGTGACCT GCTGGAGAAG
       61 ACAGACCTAG ACAGCGGAGG CTTCGGGAAG GTGTCCTTGT GTTACCACAG AAGCCATGGA
      121 TTTGTCATCC TGAAAAAAGT ATACACAGGG CCCAACCGCG CTGAGTACAA TGAGGTTCTC
      181 TTGGAAGAGG GGAAGATGAT GCACAGACTG AGACACAGTC GAGTGGTGAA GCTACTGGGC
      241 ATCATCATAG AAGAAGGGAA CTATTCGCTG GTGATGGAGT ACATGGAGAA GGGCAACCTG
      301 ATGCACGTGC TAAAGACCCA GATAGATGTC CCACTTTCAT TGAAAGGAAG GATAATCGTG
      361 GAGGCCATAG AAGGCATGTG CTACTTACAT GACAAAGGTG TGATACACAA GGACCTGAAG
      421 CCTGAGAATA TCCTCGTTGA TCGTGACTTT CACATTAAGA TAGCCGCCCT TGGTGTGGCT
      481 TCCTTTAAGA CATGGAGCAA ACTGACTAAG GAGAAAGACA ACAAGCAGAA AGAAGTGAGC
      541 AGCACCACTA AGAAGAACAA TGGTGGTACC CTTTACTACA TGGCACCCGA ACACCTGAAT
      601 GACATCAATG CAAAGCCCAC GGAGAAGTCG GACGTGTACA GCTTTGGCAT TGTCCTTTGG
      661 GCAATATTTG CAAAAAAGGA GCCCTATGAG AATGTCATCT GTACTGAGCA GTTCGTGATC
      721 TGCATAAAAT CTGGGAACAG GCCAAATGTA GAGGAAATCC TTGAGTACTG TCCAAGGGAG
      781 ATCATCAGCC TCATGGAGCG GTGCTGGCAG GCGATCCCAG AAGACAGGCC AACATTTCTT
      841 GGCATTGAAG AAGAATTTAG GCCTTTTTAC TTAAGTCATT TTGAAGAATA TGTAGAAGAG
      901 GATGTGGCAA GTTTAAAGAA AGAGTATCCA GATCAAAGCC CAGTGCTGCA GAGAATGTTT
      961 TCACTGCAGC ATGACTGTGT ACCCTTACCT CCGAGCAGGT CAAATTCAGA ACAACCTGGA
     1021 TCGCTGCACA GTTCCCAGGG GCTCCAGATG GGTCCTGTGG AGGAGTCCTG GTTTTCTTCC
     1081 TCCCCAGAGT ACCCACAGGA CGAGAATGAT CGCAGTGTGC AGGCTAAGCT GCAAGAGGAA
     1141 GCCAGCTATC ATGCTTTTGG AATATTTGCA GAGAAACAGA CAAAACCGCA GCCAAGGCAG
     1201 AATGAGGCTT ACAACAGAGA GGAGGAAAGG AAACGAAGGG TCTCTCATGA CCCCTTTGCA
     1261 CAGCAGAGAG CTCGTGAGAA TATTAAGAGT GCAGGAGCAA GAGGTCATTC TGATCCCAGC
     1321 ACAACGAGTC GTGGAATTGC AGTGCAACAG CTGTCATGGC CAGCCACCCA AACAGTTTGG
     1381 AACAATGGAT TGTATAATCA GCATGGATTT GGAACTACAG GTACAGGAGT TTGGTATCCG
     1441 CCAAATCTAA GCCAAATGTA TAGTACTTAT AAAACTCCAG TGCCAGAGAC CAACATACCG
     1501 GGAAGCACAC CCACCATGCC ATACTTCTCT GGGCCAGTAG CAGATGACCT CATAAAATAT
     1561 ACTATATTCA ATAGTTCTGG TATTCAGATT GGAAACCACA ATTATATGGA TGTTGGACTG
     1621 AATTCACAAC CACCAAACAA TACTTGCAAA GAAGAGTCGA CTTAATACCC AGCTTTCTTG
     1681 TACAAAGTGG GCATTATAAG AAAGCATTGC TTATCAATTT GTTGCAACGA ACAGGTCACT
     1741 ATCAGTCAAA ATAAAATCAT TATTTGCCAT CCAGCTGCAG CTCTGGCCCG TGTCTCAAAA
     1801 TCTCTGATGT TACATTGCAC AAGATAAAAA TATATCATCA TGAACAATAA AACTGTCTGC
     1861 TTACATAAAC AGTAATACAA GGGGTGTTAT GAGCCATATT CAACGGGAAA CGTCGAGGCC
     1921 GCGATTAAAT TCCAACATGG ATGCTGATTT ATATGGGTAT AAATGGGCTC GCGATAATGT
     1981 CGGGCAATCA GGTGCGACAA TCTATCGCTT GTATGGGAAG CCCGATGCGC CAGAGTTGTT
     2041 TCTGAAACAT GGCAAAGGTA GCGTTGCCAA TGATGTTACA GATGAGATGG TCAGACTAAA
     2101 CTGGCTGACG GAATTTATGC CTCTTCCGAC CATCAAGCAT TTTATCCGTA CTCCTGATGA
     2161 TGCATGGTTA CTCACCACTG CGATCCCCGG AAAAACAGCA TTCCAGGTAT TAGAAGAATA
     2221 TCCTGATTCA GGTGAAAATA TTGTTGATGC GCTGGCAGTG TTCCTGCGCC GGTTGCATTC
     2281 GATTCCTGTT TGTAATTGTC CTTTTAACAG CGATCGCGTA TTTCGTCTCG CTCAGGCGCA
     2341 ATCACGAATG AATAACGGTT TGGTTGATGC GAGTGATTTT GATGACGAGC GTAATGGCTG
     2401 GCCTGTTGAA CAAGTCTGGA AAGAAATGCA TAAACTTTTG CCATTCTCAC CGGATTCAGT
     2461 CGTCACTCAT GGTGATTTCT CACTTGATAA CCTTATTTTT GACGAGGGGA AATTAGGCCG
     2521 GGAAGCCGAT CTCGGCTTGA ACGAATTGTT AGGTGGCGGT ACTTGGGTCG ATATCAAAGT
     2581 GCATCACTTC TTCCCGTATG CCCAACTTTG TATAGAGAGC CACTGCGGGA TCGTCACCGT
     2641 AATCTGCTTG CACGTAGATC ACATAAGCAC CAAGCGCGTT GGCCTCATGC TTGAGGAGAT
     2701 TGATGAGCGC GGTGGCAATG CCCTGCCTCC GGTGCTCGCC GGAGACTGCG AGATCATAGA
     2761 TATAGATCTC ACTACGCGGC TGCTCAAACC TGGGCAGAAC GTAAGCCGCG AGAGCGCCAA
     2821 CAACCGCTTC TTGGTCGAAG GCAGCAAGCG CGATGAATGT CTTACTACGG AGCAAGTTCC
     2881 CGAGGTAATC GGAGTCCGGC TGATGTTGGG AGTAGGTGGC TACGTCTCCG AACTCACGAC
     2941 CGAAAAGATC AAGAGCAGCC CGCATGGATT TGACTTGGTC AGGGCCGAGC CTACATGTGC
     3001 GAATGATGCC CATACTTGAG CCACCTAACT TTGTTTTAGG GCGACTGCCC TGCTGCGTAA
     3061 CATCGTTGCT GCTGCGTAAC ATCGTTGCTG CTCCATAACA TCAAACATCG ACCCACGGCG
     3121 TAACGCGCTT GCTGCTTGGA TGCCCGAGGC ATAGACTGTA CAAAAAAACA GTCATAACAA
     3181 GCCATGAAAA CCGCCACTGC GCCGTTACCA CCGCTGCGTT CGGTCAAGGT TCTGGACCAG
     3241 TTGCGTGAGC GCATACGCTA CTTGCATTAC AGTTTACGAA CCGAACAGGC TTATGTCAAC
     3301 TGGGTTCGTG CCTTCATCCG TTTCCACGGT GTGCGTCACC CGGCAACCTT GGGCAGCAGC
     3361 GAAGTCGAGG CATTTCTGTC CTGGCTGGCG AACGAGCGCA AGGTTTCGGT CTCCACGCAT
     3421 CGTCAGGCAT TGGCGGCCTT GCTGTTCTTC TACGGCAAGG TGCTGTGCAC GGATCTGCCC
     3481 TGGCTTCAGG AGATCGGAAG ACCTCGGCCG TCGCGGCGCT TGCCGGTGGT GCTGACCCCG
     3541 GATGAAGTGG TTCGCATCCT CGGTTTTCTG GAAGGCGAGC ATCGTTTGTT CGCCCAGGAC
     3601 TCTAGCTATA GTTCTAGTGG TTGGCTACTA ATAGGTTGTA TTGATGTTGG ACGAGTCGGA
     3661 ATCGCAGACC GATACCAGGA TCTTGCCATC CTATGGAACT GCCTCGGTGA GTTTTCTCCT
     3721 TCATTACAGA AACGGCTTTT TCAAAAATAT GGTATTGATA ATCCTGATAT GAATAAATTG
     3781 CAGTTTCATT TGATGCTCGA TGAGTTTTTC TAATCAGAAT TGGTTAATTG GTTGTAACAC
     3841 TGGCAGAGCA TTACGCTGAC TTGACGGGAC GGCGCAAGCT CATGACCAAA ATCCCTTAAC
     3901 GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG
     3961 ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG
     4021 TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA
     4081 GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA
     4141 ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA
     4201 GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC
     4261 AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA
     4321 CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA
     4381 AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC
     4441 CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC
     4501 GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG
     4561 CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT
     4621 CCCCTGATTC TGTGGATAAC CGTATTACCG CTAGCCAGGA AGAGTTTGTA GAAACGCAAA
     4681 AAGGCCATCC GTCAGGATGG CCTTCTGCTT AGTTTGATGC CTGGCAGTTT ATGGCGGGCG
     4741 TCCTGCCCGC CACCCTCCGG GCCGTTGCTT CACAACGTTC AAATCCGCTC CCGGCGGATT
     4801 TGTCCTACTC AGGAGAGCGT TCACCGACAA ACAACAGATA AAACGAAAGG CCCAGTCTTC
     4861 CGACTGAGCC TTTCGTTTTA TTTGATGCCT GGCAGTTCCC TACTCTCGCG TTAACGCTAG
     4921 CATGGATCTC GGGCCCCAAA TAATGATTTT ATTTTGACTG ATAGTGACCT GTTCGTTGCA
     4981 ACAAATTGAT GAGCAATGCT TTTTTATAAT GCCAAGTTTG TACAAAAAAG CAGGCTTACT
     5041 GGAAGTTCTG TTCCAGGGGC CC
//
