LOCUS       pENTR3C-strep-hMLKL(211-471)-S358A-stop      3111 bp    DNA     circular SYN 26-FEB-2013
DEFINITION  LMBP 8708.


COMMENT     VNTOAUTHORNAME|BCCM/GeneCorner Plasmid Collection|
COMMENT     VNTOAUTHORTEL|0032-9-33-13-843|
COMMENT     VNTOAUTHOREML|bccm.genecorner@UGent.be|
COMMENT     VNTOAUTHORWWW|www.genecorner.ugent.be|
COMMENT     VNTOAUTHORAD1|Ghent University|
COMMENT     VNTOAUTHORAD2|Technologiepark 927|
COMMENT     VNTOAUTHORAD3|B-9052 Gent-Zwijnaarde Belgium|

FEATURES             Location/Qualifiers
     Source          join(<1327..3111,1..>496)
                     /label=Source_1
     Vector          join(1327..3111,1..496)
                     /origin="pENTR3C (2.723bp circular)Target vector: fromXhoI
                     cut at 938^939 to EcoRI cut at 496^497,2.281bpOrientation:
                     originalPlus strand 5':   nochangePlus strand 3':   no
                     changeMinus strand 5':  nochangeMinus strand 3':  no
                     change"
     Terminator      complement(106..149)
                     /label="rrnB T1 transcription terminator"
                     /label="rrnB T1 transcription terminator"
     Terminator      complement(281..308)
                     /label="rrnB T2 transcription terminator"
                     /label="rrnB T2 transcription terminator"
     Primer_binding_ 301..320
                     /label="pENTR FWD"
                     /label="pENTR FWD"
     Misc._recombina 358..457
                     /label=attL1
     PCR_fragment    join(<497..567,568..1298,complement(1299..>1330))
                     /note="Forward primer name: YD261-FWhMLKL+strep+C-term(EZ)"
                     /note="Forward
                     primer:AtcCggtaccgaattcAccatGgctagcTggaGccaCCCGcaGtTcgaaaaA
                     gccGAAAATGAAGTCAGCACACTTTATAAA"
                     /note="Forward primer misprimes: 42"
                     /note="Reverse primer name: YD223-RV hMLKL-stop(EZ)"
                     /note="Reverse
                     primer:gtctagatatctcgagTgCTTAGAAAAGGTGGAGAGTTTCTTT"
                     /note="Reverse primer misprimes: 17"
                     /note="Fragment length: 856"
                     /label="Fragment (hMLKL (YD261-FW hMLKL+strep+C-term(EZ) -
                     YD223-RV hMLKL-stop(EZ)))"
                     /label="Fragment (hMLKL (YD261-FW hMLKL+strep+C-term(EZ) - YD223-RV hMLKL-stop(EZ)))"
     CDS             505..540
                     /label=strep
     CDS             541..1323
                     /label="hMLKL 211-471-stop"
                     /label="hMLKL(211-471) - S358A - stop"
     Misc._recombina complement(1340..1439)
                     /label=attL2
     Primer_binding_ complement(1557..1577)
                     /label="pENTR RV"
                     /label="pENTR RV"
     CDS             1562..2371
                     /label=Kan(R)
     Replication_ori 2435..3108
                     /label="pUC origin"
                     /label="pUC origin"
ORIGIN
        1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCTAGCA TGGATCTCGG
       61 GGACGTCTAA CTACTAAGCG AGAGTAGGGA ACTGCCAGGC ATCAAATAAA ACGAAAGGCT
      121 CAGTCGGAAG ACTGGGCCTT TCGTTTTATC TGTTGTTTGT CGGTGAACGC TCTCCTGAGT
      181 AGGACAAATC CGCCGGGAGC GGATTTGAAC GTTGTGAAGC AACGGCCCGG AGGGTGGCGG
      241 GCAGGACGCC CGCCATAAAC TGCCAGGCAT CAAACTAAGC AGAAGGCCAT CCTGACGGAT
      301 GGCCTTTTTG CGTTTCTACA AACTCTTCCT GTTAGTTAGT TACTTAAGCT CGGGCCCCAA
      361 ATAATGATTT TATTTTGACT GATAGTGACC TGTTCGTTGC AACAAATTGA TAAGCAATGC
      421 TTTTTTATAA TGCCAACTTT GTACAAAAAA GCAGGCTCTT TAAAGGAACC AATTCAGTCG
      481 ACTGGATCCG GTACCGAATT CACCATGGCT AGCTGGAGCC ACCCGCAGTT CGAAAAAGCC
      541 GAAAATGAAG TCAGCACACT TTATAAAGGA GAATACCACA GAGCTCCAGT GGCCATAAAA
      601 GTATTCAAAA AACTCCAGGC TGGCAGCATT GCAATAGTGA GGCAGACTTT CAATAAGGAG
      661 ATCAAAACCA TGAAGAAATT CGAATCTCCC AACATCCTGC GTATATTTGG GATTTGCATT
      721 GATGAAACAG TGACTCCGCC TCAATTCTCC ATTGTCATGG AGTACTGTGA ACTCGGGACC
      781 CTGAGGGAGC TGTTGGATAG GGAAAAAGAC CTCACACTTG GCAAGCGCAT GGTCCTAGTC
      841 CTGGGGGCAG CCCGAGGCCT ATACCGGCTA CACCATTCAG AAGCACCTGA ACTCCACGGA
      901 AAAATCAGAA GCTCAAACTT CCTGGTAACT CAAGGCTACC AAGTGAAGCT TGCAGGATTT
      961 GAGTTGAGGA AAACACAGAC TGCCATGAGT TTGGGAACTA CGAGAGAAAA GACAGACAGA
     1021 GTCAAATCTA CAGCATATCT CTCACCTCAG GAACTGGAAG ATGTATTTTA TCAATATGAT
     1081 GTAAAGTCTG AAATATACAG CTTTGGAATC GTCCTCTGGG AAATCGCCAC TGGAGATATC
     1141 CCGTTTCAAG GCTGTAATTC TGAGAAGATC CGCAAGCTGG TGGCTGTGAA GCGGCAGCAG
     1201 GAGCCACTGG GTGAAGACTG CCCTTCAGAG CTGCGGGAGA TCATTGATGA GTGCCGGGCC
     1261 CATGATCCCT CTGTGCGGCC CTCTGTGGAT GAAATCTTAA AGAAACTCTC CACCTTTTCT
     1321 AAGCACTCGA GATATCTAGA CCCAGCTTTC TTGTACAAAG TTGGCATTAT AAGAAAGCAT
     1381 TGCTTATCAA TTTGTTGCAA CGAACAGGTC ACTATCAGTC AAAATAAAAT CATTATTTGC
     1441 CATCCAGCTG CAGCTCTGGC CCGTGTCTCA AAATCTCTGA TGTTACATTG CACAAGATAA
     1501 AAATATATCA TCATGAACAA TAAAACTGTC TGCTTACATA AACAGTAATA CAAGGGGTGT
     1561 TATGAGCCAT ATTCAACGGG AAACGTCGAG GCCGCGATTA AATTCCAACA TGGATGCTGA
     1621 TTTATATGGG TATAAATGGG CTCGCGATAA TGTCGGGCAA TCAGGTGCGA CAATCTATCG
     1681 CTTGTATGGG AAGCCCGATG CGCCAGAGTT GTTTCTGAAA CATGGCAAAG GTAGCGTTGC
     1741 CAATGATGTT ACAGATGAGA TGGTCAGACT AAACTGGCTG ACGGAATTTA TGCCTCTTCC
     1801 GACCATCAAG CATTTTATCC GTACTCCTGA TGATGCATGG TTACTCACCA CTGCGATCCC
     1861 CGGAAAAACA GCATTCCAGG TATTAGAAGA ATATCCTGAT TCAGGTGAAA ATATTGTTGA
     1921 TGCGCTGGCA GTGTTCCTGC GCCGGTTGCA TTCGATTCCT GTTTGTAATT GTCCTTTTAA
     1981 CAGCGATCGC GTATTTCGTC TCGCTCAGGC GCAATCACGA ATGAATAACG GTTTGGTTGA
     2041 TGCGAGTGAT TTTGATGACG AGCGTAATGG CTGGCCTGTT GAACAAGTCT GGAAAGAAAT
     2101 GCATAAACTT TTGCCATTCT CACCGGATTC AGTCGTCACT CATGGTGATT TCTCACTTGA
     2161 TAACCTTATT TTTGACGAGG GGAAATTAAT AGGTTGTATT GATGTTGGAC GAGTCGGAAT
     2221 CGCAGACCGA TACCAGGATC TTGCCATCCT ATGGAACTGC CTCGGTGAGT TTTCTCCTTC
     2281 ATTACAGAAA CGGCTTTTTC AAAAATATGG TATTGATAAT CCTGATATGA ATAAATTGCA
     2341 GTTTCATTTG ATGCTCGATG AGTTTTTCTA ATCAGAATTG GTTAATTGGT TGTAACATTA
     2401 TTCAGATTGG GCCCCGTTCC ACTGAGCGTC AGACCCCGTA GAAAAGATCA AAGGATCTTC
     2461 TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC
     2521 AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT
     2581 CAGCAGAGCG CAGATACCAA ATACTGTTCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT
     2641 CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC
     2701 TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA
     2761 GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC
     2821 CTACACCGAA CTGAGATACC TACAGCGTGA GCTATGAGAA AGCGCCACGC TTCCCGAAGG
     2881 GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA
     2941 GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT
     3001 TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA
     3061 CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT GCTCACATGT T
//
