LOCUS       pET-PE2-His_(IK1822)       11619 bp    DNA     circular UNA 06-APR-2021
DEFINITION  T7 promoter bacterial expression plasmid for PE2
            (nSpCas9(H840A)-MMLV-RT) with C-terminal 6xHis-tag and bipartite
            NLS.
ACCESSION   urn.local...3c5-d7ukzce
VERSION     urn.local...3c5-d7ukzce
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     rep_origin      12..467
                     /direction="RIGHT"
                     /note="arrow  indicates direction of (+) strand synthesis;
                     f1 bacteriophage origin of replication"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="f1 ori"
     primer_bind     complement(99..118)
                     /note="F1 origin, reverse primer"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="F1ori-R"
     primer_bind     309..330
                     /note="F1 origin, forward primer"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="F1ori-F"
     cds             complement(559..1374)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="confers resistance to kanamycin in bacteria or G418
                      (Geneticin(R)) in eukaryotes"
                     /Transferred_Translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDN
                     VGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRT
                     PDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFR
                     LAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIF
                     DEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFH
                     LMLDEFF"
                     /Color="#000000"
                     /modified_by="User"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="KanR"
     primer_bind     1282..1301
                     /note="Kanamycin resistance gene, reverse primer"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="Kan-R"
     primer_bind     1366..1385
                     /note="pENTR vectors, reverse primer"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="pENTR-R"
     rep_origin      1496..2084
                     /direction="LEFT; RIGHT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="ori"
     primer_bind     1985..2004
                     /note="pBR322 origin, forward primer"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="pBR322ori-F"
     primer_bind     2238..2255
                     /note="L4440 vector, forward primer"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="L4440"
     misc_feature    2270..2409
                     /note="basis of mobility region from pBR322"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="bom"
     primer_bind     complement(2335..2354)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="pRS-marker"
     primer_bind     2495..2517
                     /note="pGEX vectors, reverse primer"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="pGEX 3'"
     cds             complement(2511..2702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low
                     copy number"
                     /Transferred_Translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDAD
                     EQADICESLHDHADELYRSCLARFGDDGENL"
                     /Color="#ff0000"
                     /modified_by="User"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="rop"
     cds             complement(3511..4593)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /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)."
                     /Transferred_Translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSA
                     KTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQL
                     GASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFL
                     DVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLT
                     RNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRV
                     GADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="lacI"
     primer_bind     4555..4574
                     /note="LacI, reverse primer"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="LacI-R"
     promoter        complement(4594..4671)
                     /gene="lacI"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="lacI promoter"
     primer_bind     4877..4896
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
                     /Transferred_From="BE3 - pET backbone, pBR322 ori-F + rop,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="pBRrevBam"
     promoter        4980..4998
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /Transferred_From="BE3 - pET backbone"
                     /Transferred_Similarity="100.00%"
                     /label="T7 promoter"
     regulatory      4999..5023
                     /created_by="User"
                     /Transferred_From="pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS
                     (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="lac operator"
     misc_feature    5026..5031
                     /Recognition_pattern="T^CTAGA"
                     /annotation_group="TCTAGA"
                     /label="XbaI"
                     /note="Geneious type: restriction site"
     rbs             5038..5060
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="RBS"
     ORF             5067..11447
                     /Genetic_code="Standard"
                     /Reading_frame=1
                     /created_by="ORF Finder"
                     /label="ORF 1 (frame 1)"
     misc_RNA        5070..5102
                     /created_by="Joung Lab"
                     /X-Transferred_From="CMV-BE4-max; ORF;
                     pCAG-Cas12i-BE4-max"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="*** pCAG-ABEmax-UNG, ***
                     pCAG-ABEmax-hUNG, *** pCAG-UNG-ABEmax, ***
                     pCAG-hUNG-ABEmax, CMV-BE4-max, pCMV_ABEmax"
                     /Transferred_Similarity="100.00%"
                     /label="bipartite NLS"
     CDS             5103..5123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     CDS             5124..9224
                     /created_by="User"
                     /modified_by="Joung Lab"
                     /label="bacteria codon optimized H840A mutant SpCas9"
     primer_bind     7638..7640
                     /created_by="Joung Lab"
                     /Sequence="GCA"
                     /Color="#ff0000"
                     /modified_by="Joung Lab"
                     /label="H840A"
     misc_RNA        9225..9323
                     /created_by="Joung Lab"
                     /X-Transferred_From="CMV-BE4-max;
                     pCMV(Δ7)-Gam-UNG-BE4max-Δ(R33A)"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="CMV-BE4-max,
                     pCMV(Δ7)-Gam-UNG-BE4max-Δ(R33A),
                     pCMV(Δ7)-UNG-deadBE4max-Δ(R33A)"
                     /Transferred_Similarity="100.00%"
                     /label="linker"
     gene            9324..11354
                     /created_by="Joung Lab"
                     /label="M-MLV RT"
     misc_feature    10742..10747
                     /Recognition_pattern="A^CCGGT"
                     /annotation_group="ACCGGT"
                     /label="AgeI"
                     /note="Geneious type: restriction site"
     misc_RNA        11355..11366
                     /created_by="Joung Lab"
                     /X-Transferred_From="CMV-BE4-max; ORF;
                     pCAG-Cas12i-BE4-max"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="*** pCAG-ABEmax-UNG, ***
                     pCAG-ABEmax-hUNG, *** pCAG-UNG-ABEmax, ***
                     pCAG-hUNG-ABEmax, CMV-BE4-max, pCMV_ABEmax"
                     /Transferred_Similarity="100.00%"
                     /label="linker"
     primer_bind     11367..11417
                     /Sequence="AAAAGAACCGCCGACGGCAGCGAATTCGAGCCCAAGAAGAAGAGGAA
                     AGTC"
                     /Color="#33bbff"
                     /created_by="User"
                     /modified_by="User"
                     /X-Transferred_From="SECURE miniABEmax-V82G (pJUL1828);
                     SpACE-hUNG-Gam; addgene 122607; addgene 122610; addgene
                     122611; addgene 122611  dCas9; addgene 125612; addgene
                     125615; addgene 125616; addgene 125616  dCas9;
                     ecUNG-evoFERNY; pCMV-BE4max-UNG-Gam;
                     pCMV-Gam-miniABEmax(V82G)-UNG;
                     pCMV-NLS-rAPOBEC1-SaKKH_nCas9-P2A-GFP"
                     /X-Transferred_Similarity="100.00%"
                     /created_by="User"
                     /Transferred_From="SECURE miniABEmax-V82G (pJUL1828) ,
                     addgene 122607 , addgene 122611  dCas9, addgene 125612 ,
                     addgene 125615 , addgene 125616  dCas9, ecUNG-evoFERNY,
                     pCMV-BE4max(R33A)-UNG-Gam (T7 promoter removed),
                     pCMV-BE4max-UNG-Gam, pCMV-Gam-miniABEmax(V82G)-UNG,
                     pCMV-NLS-rAPOBEC1-SaKKH_nCas9-P2A-GFP,
                     pCMV-NLS-rAPOBEC1-SaKKH_nCas9-UNG-P2A-GFP"
                     /Transferred_Similarity="100.00%"
                     /label="bpNLS"
     CDS             11397..11417
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     polylinker      11418..11426
                     /modified_by="Joung Lab"
                     /label="linker"
     gene            11427..11444
                     /modified_by="Joung Lab"
                     /label="His-tag"
     misc_feature    11448..11455
                     /Recognition_pattern="GC^GGCCGC"
                     /annotation_group="GCGGCCGC"
                     /label="NotI"
                     /note="Geneious type: restriction site"
     terminator      11547..11594
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /Transferred_From="BE3 - pET backbone,
                     pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS (KAN)"
                     /Transferred_Similarity="100.00%"
                     /label="T7 terminator"
ORIGIN      
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      601 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      661 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      721 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      781 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      841 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      901 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      961 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1021 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     1081 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1141 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1201 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1261 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1321 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     1381 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1621 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     2281 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     4981 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     5041 gtttaacttt aagaaggaga tataccatga aacggacagc cgacggaagc gagttcgagt
     5101 caccaaagaa gaagcggaaa gtcgataaga aatactcaat aggcttagat atcggcacaa
     5161 atagcgtcgg atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg
     5221 ttctgggaaa tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg
     5281 acagtggaga gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac
     5341 gtcggaagaa tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag
     5401 atgatagttt ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg
     5461 aacgtcatcc tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa
     5521 ctatctatca tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa
     5581 tctatttggc cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt
     5641 taaatcctga taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc
     5701 aattatttga agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg
     5761 cacgattgag taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga
     5821 aaaatggctt atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat
     5881 caaattttga tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg
     5941 atttagataa tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta
     6001 agaatttatc agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta
     6061 aggctcccct atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc
     6121 ttttaaaagc tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc
     6181 aatcaaaaaa cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata
     6241 aatttatcaa accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa
     6301 atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa
     6361 ttcacttggg tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa
     6421 aagacaatcg tgagaagatt gaaaaaatct tgacttttcg aattccttat tatgttggtc
     6481 cattggcgcg tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta
     6541 ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac
     6601 gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc
     6661 tttatgagta ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa
     6721 tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca
     6781 aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat
     6841 gttttgatag tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct
     6901 accatgattt gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag
     6961 atatcttaga ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg
     7021 aaagacttaa aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc
     7081 gccgttatac tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc
     7141 aatctggcaa aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta
     7201 tgcagctgat ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt
     7261 ctggacaagg cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta
     7321 aaaaaggtat tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc
     7381 ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc
     7441 agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc
     7501 agattcttaa agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct
     7561 attatctcca aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa
     7621 gtgattatga tgtcgatgca attgttccac aaagtttcct taaagacgat tcaatagaca
     7681 ataaggtctt aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag
     7741 aagtagtcaa aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc
     7801 aacgtaagtt tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag
     7861 ctggttttat caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa
     7921 ttttggatag tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta
     7981 aagtgattac cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata
     8041 aagtacgtga gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg
     8101 gaactgcttt gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata
     8161 aagtttatga tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg
     8221 caaaatattt cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa
     8281 atggagagat tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct
     8341 gggataaagg gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata
     8401 ttgtcaagaa aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa
     8461 gaaattcgga caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt
     8521 ttgatagtcc aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat
     8581 cgaagaagtt aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct
     8641 ttgaaaaaaa tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact
     8701 taatcattaa actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc
     8761 tggctagtgc cggagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga
     8821 attttttata tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac
     8881 aaaaacaatt gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca
     8941 gtgaattttc taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat
     9001 ataacaaaca tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta
     9061 cgttgacgaa tcttggagct ccagccgcat tcaagtattt tgacacaacg atagatcgca
     9121 aacgatacac ttctaccaag gaggtgctag acgcgacact gattcaccaa tccatcacgg
     9181 gattatatga aactcggata gatttgtcac agcttggggg tgactctgga ggatctagcg
     9241 gaggatcctc tggcagcgag acaccaggaa caagcgagtc agcaacacca gagagcagtg
     9301 gcggcagcag cggcggcagc agcaccctaa atatagaaga tgagtatcgg ctacatgaga
     9361 cctcaaaaga gccagatgtt tctctagggt ccacatggct gtctgatttt cctcaggcct
     9421 gggcggaaac cgggggcatg ggactggcag ttcgccaagc tcctctgatc atacctctga
     9481 aagcaacctc tacccccgtg tccataaaac aataccccat gtcacaagaa gccagactgg
     9541 ggatcaagcc ccacatacag agactgttgg accagggaat actggtaccc tgccagtccc
     9601 cctggaacac gcccctgcta cccgttaaga aaccagggac taatgattat aggcctgtcc
     9661 aggatctgag agaagtcaac aagcgggtgg aagacatcca ccccaccgtg cccaaccctt
     9721 acaacctctt gagcgggctc ccaccgtccc accagtggta cactgtgctt gatttaaagg
     9781 atgccttttt ctgcctgaga ctccacccca ccagtcagcc tctcttcgcc tttgagtgga
     9841 gagatccaga gatgggaatc tcaggacaat tgacctggac cagactccca cagggtttca
     9901 aaaacagtcc caccctgttt aatgaggcac tgcacagaga cctagcagac ttccggatcc
     9961 agcacccaga cttgatcctg ctacagtacg tggatgactt actgctggcc gccacttctg
    10021 agctagactg ccaacaaggt actcgggccc tgttacaaac cctagggaac ctcgggtatc
    10081 gggcctcggc caagaaagcc caaatttgcc agaaacaggt caagtatctg gggtatcttc
    10141 taaaagaggg tcagagatgg ctgactgagg ccagaaaaga gactgtgatg gggcagccta
    10201 ctccgaagac ccctcgacaa ctaagggagt tcctagggaa ggcaggcttc tgtcgcctct
    10261 tcatccctgg gtttgcagaa atggcagccc ccctgtaccc tctcaccaaa ccggggactc
    10321 tgtttaattg gggcccagac caacaaaagg cctatcaaga aatcaagcaa gctcttctaa
    10381 ctgccccagc cctggggttg ccagatttga ctaagccctt tgaactcttt gtcgacgaga
    10441 agcagggcta cgccaaaggt gtcctaacgc aaaaactggg accttggcgt cggccggtgg
    10501 cctacctgtc caaaaagcta gacccagtag cagctgggtg gcccccttgc ctacggatgg
    10561 tagcagccat tgccgtactg acaaaggatg caggcaagct aaccatggga cagccactag
    10621 tcattctggc cccccatgca gtagaggcac tagtcaaaca accccccgac cgctggcttt
    10681 ccaacgcccg gatgactcac tatcaggcct tgcttttgga cacggaccgg gtccagttcg
    10741 gaccggtggt agccctgaac ccggctacgc tgctcccact gcctgaggaa gggctgcaac
    10801 acaactgcct tgatatcctg gccgaagccc acggaacccg acccgaccta acggaccagc
    10861 cgctcccaga cgccgaccac acctggtaca cggatggaag cagtctctta caagagggac
    10921 agcgtaaggc gggagctgcg gtgaccaccg agaccgaggt aatctgggct aaagccctgc
    10981 cagccgggac atccgctcag cgggctgaac tgatagcact cacccaggcc ctaaagatgg
    11041 cagaaggtaa gaagctaaat gtttatactg atagccgtta tgcttttgct actgcccata
    11101 tccatggaga aatatacaga aggcgtgggt ggctcacatc agaaggcaaa gagatcaaaa
    11161 ataaagacga gatcttggcc ctactaaaag ccctctttct gcccaaaaga cttagcataa
    11221 tccattgtcc aggacatcaa aagggacaca gcgccgaggc tagaggcaac cggatggctg
    11281 accaagcggc ccgaaaggca gccatcacag agactccaga cacctctacc ctcctcatag
    11341 aaaattcatc accctctggc ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca
    11401 agaagaagag gaaagtcggc agcagccatc atcatcatca tcactaagcg gccgcactcg
    11461 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
    11521 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
    11581 tgaggggttt tttgctgaaa ggaggaacta tatccggat
//
