pET28b-BE1

LOCUS       pET28b-BE1               10203 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Expresses BE1 with N-terminal His6 tag in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    pET28b-BE1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10203)
  AUTHORS   Komor AC, Kim YB, Packer MS, Zuris JA, Liu DR
  TITLE     Programmable editing of a target base in genomic DNA without 
            double-stranded DNA cleavage.
  JOURNAL   Nature. 2016 Apr 20. doi: 10.1038/nature17946.
  PUBMED    27096365
REFERENCE   2  (bases 1 to 10203)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10203
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(559..1374)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1282..1301
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1366..1385
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1496..2084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1985..2004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2238..2255
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2270..2409
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2335..2354)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2495..2517
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2511..2702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3511..4593)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4555..4574
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4594..4671)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4877..4896
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4980..4999
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4980..4998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4999..5023
                     /label=lac operator
                     /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)."
     RBS             5038..5060
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5079..5096
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5097..5783
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             5832..9935
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9945..9962
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             9966..9998
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             10047..10064
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(10117..10135)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      10131..10178
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
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 tataccatgg gcagcagcca tcatcatcat catcacatgt
     5101 cttctgaaac cggtccggtt gcggttgacc cgaccctgcg tcgtcgtatc gaaccgcacg
     5161 aattcgaagt tttcttcgac ccgcgtgaac tgcgtaaaga aacctgcctg ctgtacgaaa
     5221 tcaactgggg tggtcgtcac tctatctggc gtcacacctc tcagaacacc aacaaacacg
     5281 ttgaagttaa cttcatcgaa aaattcacca ccgaacgtta cttctgcccg aacacccgtt
     5341 gctctatcac ctggttcctg tcttggtctc cgtgcggtga atgctctcgt gcgatcaccg
     5401 aattcctgtc tcgttacccg cacgttaccc tgttcatcta catcgcgcgt ctgtaccacc
     5461 acgcggaccc gcgtaaccgt cagggtctgc gtgacctgat ctcttctggt gttaccatcc
     5521 agatcatgac cgaacaggaa tctggttact gctggcgtaa cttcgttaac tactctccgt
     5581 ctaacgaagc gcactggccg cgttacccgc acctgtgggt tcgtctgtac gttctggaac
     5641 tgtactgcat catcctgggt ctgccgccgt gcctgaacat cctgcgtcgt aaacagccgc
     5701 agctgacctt cttcaccatc gcgctgcagt cttgccacta ccagcgtctg ccgccgcaca
     5761 tcctgtgggc gaccggtctg aaatccggta gcgaaacacc ggggacttca gaatcggcca
     5821 ccccggagtc tgataagaaa tactcaatag gcttagctat cggcacaaat agcgtcggat
     5881 gggcggtgat cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata
     5941 cagaccgcca cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga
     6001 cagcggaagc gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc
     6061 gtatttgtta tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct
     6121 ttcatcgact tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta
     6181 tttttggaaa tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc
     6241 tgcgaaaaaa attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct
     6301 tagcgcatat gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata
     6361 atagtgatgt ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag
     6421 aaaaccctat taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta
     6481 aatcaagacg attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat
     6541 ttgggaatct cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt
     6601 tggcagaaga tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt
     6661 tattggcgca aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag
     6721 atgctatttt actttcagat atcctaagag taaatactga aataactaag gctcccctat
     6781 cagcttcaat gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt
     6841 tagttcgaca acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg
     6901 gatatgcagg ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac
     6961 caattttaga aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt
     7021 tgctgcgcaa gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg
     7081 agctgcatgc tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg
     7141 agaagattga aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg
     7201 gcaatagtcg ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt
     7261 ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacaaact
     7321 ttgataaaaa tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt
     7381 ttacggttta taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag
     7441 catttctttc aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa
     7501 aagtaaccgt taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg
     7561 ttgaaatttc aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc
     7621 taaaaattat taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg
     7681 atattgtttt aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa
     7741 catatgctca cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg
     7801 gttggggacg tttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa
     7861 caatattaga ttttttgaaa tcagatggtt ttgccaatcg caattttatg cagctgatcc
     7921 atgatgatag tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg
     7981 atagtttaca tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt
     8041 tacagactgt aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa
     8101 atatcgttat tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc
     8161 gagagcgtat gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag
     8221 agcatcctgt tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa
     8281 atggaagaga catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg
     8341 tcgatgccat tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa
     8401 cgcgttctga taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa
     8461 agatgaaaaa ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg
     8521 ataatttaac gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca
     8581 aacgccaatt ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc
     8641 gcatgaatac taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct
     8701 taaaatctaa attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga
     8761 ttaacaatta ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga
     8821 ttaagaaata tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg
     8881 ttcgtaaaat gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct
     8941 tttactctaa tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc
     9001 gcaaacgccc tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc
     9061 gagattttgc cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa
     9121 cagaagtaca gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca
     9181 agcttattgc tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa
     9241 cggtagctta ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa
     9301 aatccgttaa agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc
     9361 cgattgactt tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac
     9421 tacctaaata tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg
     9481 gagaattaca aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt
     9541 tagctagtca ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt
     9601 ttgtggagca gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta
     9661 agcgtgttat tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata
     9721 gagacaaacc aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc
     9781 ttggagctcc cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt
     9841 ctacaaaaga agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa
     9901 cacgcattga tttgagtcag ctaggaggtg actgaagcag cggcctggtg ccgcgcggca
     9961 gccatatggc tagcatgact ggtggacagc aaatgggtcg ggatccgaat tcgagctccg
    10021 tcgacaagct tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta
    10081 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
    10141 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
    10201 gat
//