LOCUS       PD0028_pIVTRup(BsmBI)_dCas9        8463 bp    DNA     circular UNA 07-JAN-2020
DEFINITION  Concatenation of 2 sequences.
ACCESSION   urn.local...4z-bsz22p6
VERSION     urn.local...4z-bsz22p6
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     origin          90^91
                     /label="origin"
     promoter        165..183
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     235..295
                     /Sequence="TTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAG
                     AGAGAAAAGAAGAG"
                     /Mismatches=0
                     /annotation_group="a24e0d84-0d69-45ce-a2d0-cea8b5c33ed8"
                     /label="pIVTRup-BsmBI-FWD"
     primer_bind     235..295
                     /Sequence="TTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAG
                     AGAGAAAAGAAGAG"
                     /created_by="User"
                     /label="pIVTRup-FWD-AG"
     promoter        255..273
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     cds             272..>301
                     /created_by="User"
                     /modified_by="User"
                     /X-Transferred_From="101362 - pIVTRup Copy"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="101362 - pIVTRup Copy, 101362 - pIVTRup
                     PCR DraI, Digestion of 101362 - pIVTRup PCR DraI -
                     Fragment 3"
                     /Transferred_Similarity="100.00%"
                     /label="3'UTR"
     primer_bind     complement(293..318)
                     /Sequence="GGTGGCTCTTATATTTCTTCTTACTC"
                     /Hairpin_Tm="None"
                     /Tm="57.4"
                     /Self_Dimer_Tm="None"
                     /%GC="38.5"
                     /modified_by="User"
                     /label="pIVTRup_linearize_REV"
     misc_feature    301^302
                     /Original_Bases="GAAATATAAGAGCCACCGAT"
                     /label="GAAATATAAGAGCCACCGAT"
                     /note="Geneious type: Editing History Deletion"
     primer_bind     322..342
                     /Mismatches=0
                     /Extension="GAAATATAAGAGCCACCGAT"
                     /created_by="primer3"
                     /annotation_group="24245091615550df09c719-883f-44d0-9e59-d
                     be6cb4b677c"
                     /%GC="66.7"
                     /Tm="68.7"
                     /Hairpin_Tm="68.2"
                     /Self_Dimer_Tm="56.1"
                     /Sequence="ATGAAAAGGCCGGCGGCCACG"
                     /label="dCas9_FWD"
     CDS             325..372
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /translation="KRPAATKKAGQAKKKK"
                     /label="nucleoplasmin NLS"
     CDS             373..4473
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system, mutated to improve
                     targeting specificity (Slaymaker et al., 2016)"
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEE
                     SFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHM
                     IKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKS
                     RRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDN
                     LLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLL
                     KALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKL
                     NREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYY
                     VGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLP
                     KHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKED
                     YFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTL
                     FEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFL
                     KSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTV
                     KVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEH
                     PVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVL
                     TRSDKARGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAG
                     FIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFY
                     KVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
                     ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSM
                     PQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVA
                     KVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFE
                     LENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHK
                     HYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAP
                     AAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
                     /label="eSpCas9(1.1)"
     primer_bind     720..739
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="65416116108827cd833bd-0bf5-4433-abb2-0e
                     eda78a8d49"
                     /%GC="55.0"
                     /Tm="59.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="CATCTTCGGCAACATCGTGG"
                     /label="PCR_0268"
     primer_bind     1292..1312
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="6541662943741f426e091-ff62-4b70-96f2-f3
                     1190877804"
                     /%GC="52.4"
                     /Tm="60.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="TGAACACCGAGATCACCAAGG"
                     /label="PCR_0269"
     primer_bind     1771..1791
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="654170983114734c3bf80-f4fa-4675-9a6f-2f
                     76f160724f"
                     /%GC="52.4"
                     /Tm="60.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="AAGAGCGAGGAAACCATCACC"
                     /label="PCR_0270"
     primer_bind     2231..2251
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="6541754887948f78d9012-a6d5-4f07-937e-92
                     349869d10e"
                     /%GC="52.4"
                     /Tm="59.9"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="TGACCCTGACACTGTTTGAGG"
                     /label="PCR_0271"
     primer_bind     2723..2743
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="6541815932605dc803015-44e3-4c83-89ab-89
                     3e4ddb5482"
                     /%GC="52.4"
                     /Tm="60.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="0.6"
                     /Sequence="AAGAGGGCATCAAAGAGCTGG"
                     /label="PCR_0272"
     primer_bind     3233..3253
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="6541856091642cf39fdf0-afd7-47dd-a708-68
                     1475b3d6d8"
                     /%GC="52.4"
                     /Tm="60.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="TGATCACCCTGAAGTCCAAGC"
                     /label="PCR_0273"
     primer_bind     3842..3862
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="65418941624069c830598-e6c5-4566-8101-a0
                     6644791a6f"
                     /%GC="52.4"
                     /Tm="60.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="7.0"
                     /Sequence="AGAGTGTGAAAGAGCTGCTGG"
                     /label="PCR_0274"
     primer_bind     4280..4300
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="6541972521111799a435e-0b82-4cb0-b2ad-2a
                     5dce3ffb0d"
                     /%GC="52.4"
                     /Tm="59.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="AGCAGGCCGAGAATATCATCC"
                     /label="PCR_0275"
     CDS             4528..4548
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     misc_feature    complement(4553..4572)
                     /Mismatches=0
                     /Extension="GCCCCGCAGAAGGCAGCGATTTA"
                     /created_by="primer3"
                     /annotation_group="242451483951913212becd-95df-4205-a123-c
                     d02f57a3378"
                     /%GC="75.0"
                     /Tm="69.0"
                     /Hairpin_Tm="41.8"
                     /Self_Dimer_Tm="23.8"
                     /Sequence="AGCCCGTCCGGATCCAGCGG"
                     /label="dCas9_REV"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     4576..4591
                     /Sequence="ATCGCTGCCTTCTGCG"
                     /Hairpin_Tm="53.8"
                     /Tm="57.2"
                     /Self_Dimer_Tm="9.7"
                     /%GC="62.5"
                     /created_by="primer3"
                     /label="pIVTRup_linearize_FWD"
     misc_feature    4595^4596
                     /Original_Bases="ATCGCTGCCTTCTGCGGGGC"
                     /label="ATCGCTGCCTTCTGCGGGGC"
                     /note="Geneious type: Editing History Deletion"
     cds             <4596..4682
                     /created_by="User"
                     /modified_by="User"
                     /X-Transferred_From="101362 - pIVTRup Copy; dCas9-VP64"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="101362 - pIVTRup Copy, 101362 - pIVTRup
                     PCR DraI, Digestion of 101362 - pIVTRup PCR DraI -
                     Fragment 3, dCas9-VP64"
                     /Transferred_Similarity="100.00%"
                     /label="5'UTR"
     misc_feature    complement(4667..4737)
                     /Sequence="TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
                     TTTTTTTTCTAGACCCTCACTTCC"
                     /Mismatches=0
                     /annotation_group="134d99b0-360b-4f39-8356-3ca8ae430d72"
                     /label="pIVTRup-BsmBI-REV"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     complement(4667..4737)
                     /Sequence="TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
                     TTTTTTTTCTAGACCCTCACTTCC"
                     /created_by="User"
                     /label="pIVTRup-REV"
     3'UTR           4683..4737
                     /created_by="User"
                     /label="polyA(55)"
     misc_feature    complement(4739..4744)
                     /Recognition_pattern="CGTCTC(1/5)"
                     /annotation_group="CGTCTC"
                     /label="BsmBI"
                     /note="Geneious type: restriction site"
     protein_bind    4883..4899
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4907..4937)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4952..4973
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5261..5849)
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     rep_origin      complement(8036..1)
                     /direction="LEFT"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN      
        1 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
      181 agggcgaatt gggccctcta gatgcatgct cgagcggccg ccagtgtgat ggatttggac
      241 cctcgtacag aagctaatac gactcactat agggaaataa gagagaaaag aagagtaaga
      301 agaaatataa gagccaccga tatgaaaagg ccggcggcca cgaaaaaggc cggccaggca
      361 aaaaagaaaa aggacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
      421 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
      481 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
      541 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
      601 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
      661 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
      721 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
      781 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
      841 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
      901 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
      961 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     1021 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     1081 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     1141 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     1201 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     1261 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     1321 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     1381 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     1441 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     1501 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     1561 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     1621 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     1681 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     1741 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     1801 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     1861 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     1921 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     1981 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     2041 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     2101 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     2161 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     2221 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     2281 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     2341 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     2401 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     2461 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     2521 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     2581 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     2641 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     2701 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     2761 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     2821 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     2881 gtggaccaca tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     2941 accagaagcg acaaggcccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     3001 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     3061 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     3121 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     3181 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     3241 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     3301 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     3361 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     3421 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     3481 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     3541 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     3601 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     3661 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     3721 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     3781 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     3841 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     3901 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     3961 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     4021 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     4081 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     4141 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     4201 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     4261 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     4321 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     4381 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     4441 acacggatcg acctgtctca gctgggaggc gacagcgctg gaggaggtgg aagcggagga
     4501 ggaggaagcg gaggaggagg tagcggacct aagaaaaaga ggaaggtggc ggccgctgga
     4561 tccggacggg cttaaatcgc tgccttctgc ggggcttgcc ttctggccat gcccttcttc
     4621 tctcccttgc acctgtacct cttggtcttt gaataaagcc tgagtaggaa gtgagggtct
     4681 agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatga
     4741 gacgcgatca ataaccgaca ctcctgcgtc gcctacgcgc tacgatctgc agaattccag
     4801 cacactggcg gccgttacta gtggatccga gctcggtacc aagcttggcg taatcatggt
     4861 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4921 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4981 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5041 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5101 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5161 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5221 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5281 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5341 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5401 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5461 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5521 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5581 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5641 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5701 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5761 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5821 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5881 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5941 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6001 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6061 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6121 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6181 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6241 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6301 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6361 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6421 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6481 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6541 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6601 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6661 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6721 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6781 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6841 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaattca
     6901 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     6961 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     7021 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     7081 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     7141 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     7201 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     7261 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     7321 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtggga
     7381 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     7441 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     7501 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     7561 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     7621 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     7681 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg
     7741 ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt
     7801 accagagggc gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc
     7861 tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc
     7921 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     7981 ggctttctac gtgttccgct tcctttagca gcccttgcgc cctgaatttt gttaaaattc
     8041 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     8101 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     8161 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     8221 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     8281 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     8341 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     8401 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     8461 gcg
//
