LOCUS       sequence-354242-       11594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing PE2 protein in E.coli for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11594)
  AUTHORS   Li H, Busquets O, Verma Y, Syed KM, Kutnowski N, Pangilinan GR, 
            Gilbert LA, Bateup HS, Rio DC, Hockemeyer D, Soldner F
  TITLE     Highly efficient generation of isogenic pluripotent stem cell models
            using prime editing.
  JOURNAL   Elife. 2022 Sep 7;11:e79208. doi: 10.7554/eLife.79208.
  PUBMED    36069759
REFERENCE   2  (bases 1 to 11594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2022-09-7"; volume: "11:e79208. doi"; pages: " 
            10.7554/eLife.79208"
COMMENT     Sequence Label: pET30a(+)-PE2
FEATURES             Location/Qualifiers
     source          1..11594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(173..193)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(2366..3949,3950..3952,3953..6466))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(2366..6466)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(6467..6487)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             6531..6553
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6568..6592
                     /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)."
     primer_bind     complement(6592..6611)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6593..6611)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6699..6718)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6924..7001
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7002..8084
                     /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     complement(7021..7040)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             8893..9084
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(9078..9100)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9186..9325
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9241..9260
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(9340..9357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9511..10099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9591..9610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(10210..10229)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10221..11036
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             10221..11036
                     /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     complement(10294..10313)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(11128..11583)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11265..11286)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11477..11496
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcgactttcc
      181 tcttcttctt gggctcgaat tcgctgccgt cggcggttct ttttgagccg ccagagggtg
      241 atgaattttc tatgaggagg gtagaggtgt ctggagtctc tgtgatggct gcctttcggg
      301 ccgcttggtc agccatccgg ttgcctctag cctcggcgct gtgtcccttt tgatgtcctg
      361 gacaatggat tatgctaagt cttttgggca gaaagagggc ttttagtagg gccaagatct
      421 cgtctttatt tttgatctct ttgccttctg atgtgagcca cccacgcctt ctgtatattt
      481 ctccatggat atgggcagta gcaaaagcat aacggctatc agtataaaca tttagcttct
      541 taccttctgc catctttagg gcctgggtga gtgctatcag ttcagcccgc tgagcggatg
      601 tcccggctgg cagggcttta gcccagatta cctcggtctc ggtggtcacc gcagctcccg
      661 ccttacgctg tccctcttgt aagagactgc ttccatccgt gtaccaggtg tggtcggcgt
      721 ctgggagcgg ctggtccgtt aggtcgggtc gggttccgtg ggcttcggcc aggatatcaa
      781 ggcagttgtg ttgcagccct tcctcaggca gtgggagcag cgtagccggg ttcagggcta
      841 ccaccggtcc gaactggacc cggtccgtgt ccaaaagcaa ggcctgatag tgagtcatcc
      901 gggcgttgga aagccagcgg tcggggggtt gtttgactag tgcctctact gcatgggggg
      961 ccagaatgac tagtggctgt cccatggtta gcttgcctgc atcctttgtc agtacggcaa
     1021 tggctgctac catccgtagg caagggggcc acccagctgc tactgggtct agctttttgg
     1081 acaggtaggc caccggccga cgccaaggtc ccagtttttg cgttaggaca cctttggcgt
     1141 agccctgctt ctcgtcgaca aagagttcaa agggcttagt caaatctggc aaccccaggg
     1201 ctggggcagt tagaagagct tgcttgattt cttgataggc cttttgttgg tctgggcccc
     1261 aattaaacag agtccccggt ttggtgagag ggtacagggg ggctgccatt tctgcaaacc
     1321 cagggatgaa gaggcgacag aagcctgcct tccctaggaa ctcccttagt tgtcgagggg
     1381 tcttcggagt aggctgcccc atcacagtct cttttctggc ctcagtcagc catctctgac
     1441 cctcttttag aagatacccc agatacttga cctgtttctg gcaaatttgg gctttcttgg
     1501 ccgaggcccg atacccgagg ttccctaggg tttgtaacag ggcccgagta ccttgttggc
     1561 agtctagctc agaagtggcg gccagcagta agtcatccac gtactgtagc aggatcaagt
     1621 ctgggtgctg gatccggaag tctgctaggt ctctgtgcag tgcctcatta aacagggtgg
     1681 gactgttttt gaaaccctgt gggagtctgg tccaggtcaa ttgtcctgag attcccatct
     1741 ctggatctct ccactcaaag gcgaagagag gctgactggt ggggtggagt ctcaggcaga
     1801 aaaaggcatc ctttaaatca agcacagtgt accactggtg ggacggtggg agcccgctca
     1861 agaggttgta agggttgggc acggtggggt ggatgtcttc cacccgcttg ttgacttctc
     1921 tcagatcctg gacaggccta taatcattag tccctggttt cttaacgggt agcaggggcg
     1981 tgttccaggg ggactggcag ggtaccagta ttccctggtc caacagtctc tgtatgtggg
     2041 gcttgatccc cagtctggct tcttgtgaca tggggtattg ttttatggac acgggggtag
     2101 aggttgcttt cagaggtatg atcagaggag cttggcgaac tgccagtccc atgcccccgg
     2161 tttccgccca ggcctgagga aaatcagaca gccatgtgga ccctagagaa acatctggct
     2221 cttttgaggt ctcatgtagc cgatactcat cttctatatt tagggtgctg ctgccgccgc
     2281 tgctgccgcc actgctctct ggtgttgctg actcgcttgt tcctggtgtc tcgctgccag
     2341 aggatcctcc gctagatcct ccagagtcac ctcccagctg agacaggtcg atccgtgtct
     2401 cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc cagcacctct ttggtgctgg
     2461 tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat
     2521 tggtcagggt aaacaggtgg atgatattct cggcctgctc tctgatgggc ttatcccggt
     2581 gcttgttgta ggcggacagc actttgtcca gattagcgtc ggccaggatc actctcttgg
     2641 agaactcgct gatctgctcg atgatctcgt ccaggtagtg cttgtgctgt tccacaaaca
     2701 gctgtttctg ctcattatcc tcgggggagc ccttcagctt ctcatagtgg ctggccaggt
     2761 acaggaagtt cacatatttg gagggcaggg ccagttcgtt tcccttctgc agttcgccgg
     2821 cagaggccag cattctcttc cggccgtttt ccagctcgaa cagggagtac ttaggcagct
     2881 tgatgatcag gtcctttttc acttctttgt agcccttggc ttccagaaag tcgatgggat
     2941 tcttctcgaa gctgcttctt tccatgatgg tgatccccag cagctctttc acactcttca
     3001 gtttcttgga cttgcccttt tccactttgg ccaccaccag cacagaatag gccacggtgg
     3061 ggctgtcgaa gccgccgtac ttcttagggt cccagtcctt ctttctggcg atcagcttat
     3121 cgctgttcct cttgggcagg atagactctt tgctgaagcc gcctgtctgc acctcggtct
     3181 ttttcacgat attcacttgg ggcatgctca gcactttccg cacggtggca aaatcccggc
     3241 ccttatccca cacgatctcc ccggtttcgc cgtttgtctc gatcagaggc cgcttccgga
     3301 tctcgccgtt ggccagggta atctcggtct tgaaaaagtt catgatgttg ctgtagaaga
     3361 agtacttggc ggtagccttg ccgatttcct gctcgctctt ggcgatcatc ttccgcacgt
     3421 cgtacacctt gtagtcgccg tacacgaact cgctttccag cttagggtac tttttgatca
     3481 gggcggttcc cacgacggcg ttcaggtagg cgtcgtgggc gtggtggtag ttgttgatct
     3541 cgcgcacttt gtaaaactgg aaatccttcc ggaaatcgga caccagcttg gacttcaggg
     3601 tgatcacttt cacttcccgg atcagcttgt cattctcgtc gtacttagtg ttcatccggg
     3661 agtccaggat ctgtgccacg tgctttgtga tctgccgggt ttccaccagc tgtctcttga
     3721 tgaagccggc cttatccagt tcgctcaggc cgcctctctc ggccttggtc agattgtcga
     3781 actttctctg ggtaatcagc ttggcgttca gcagctgccg ccagtagttc ttcatcttct
     3841 tcacgacctc ttcggagggc acgttgtcgc tcttgccccg gttcttgtcg cttctggtca
     3901 gcaccttgtt gtcgatggag tcgtccttca gaaagctctg aggcacgata gcgtccacat
     3961 cgtagtcgga cagccggttg atgtccagtt cctggtccac gtacatatcc cgcccattct
     4021 gcaggtagta caggtacagc ttctcgttct gcagctgggt gttttccacg gggtgttctt
     4081 tcaggatctg gctgcccagc tctttgatgc cctcttcgat ccgcttcatt ctctcgcggc
     4141 tgttcttctg tcccttctgg gtggtctggt tctctctggc catttcgatc acgatgttct
     4201 cgggcttgtg ccggcccatc actttcacga gctcgtccac caccttcact gtctgcagga
     4261 tgcccttctt aatggcgggg ctgccggcca gattggcaat gtgctcgtgc aggctatcgc
     4321 cctggccgga cacctgggct ttctggatgt cctctttaaa ggtcaggctg tcgtcgtgga
     4381 tcagctgcat gaagtttctg ttggcgaagc cgtcggactt caggaaatcc aggattgtct
     4441 tgccggactg cttgtcccgg atgccgttga tcagcttccg gctcagcctg ccccagccgg
     4501 tgtatctccg ccgcttcagc tgcttcatca ctttgtcgtc gaacaggtgg gcataggttt
     4561 tcagccgttc ctcgatcatc tctctgtcct caaacagtgt cagggtcagc acgatatctt
     4621 ccagaatgtc ctcgttttcc tcattgtcca ggaagtcctt gtccttgata attttcagca
     4681 gatcgtggta tgtgcccagg gaggcgttga accgatcttc cacgccggag atttccacgg
     4741 agtcgaagca ctcgattttc ttgaagtagt cctctttcag ctgcttcacg gtcactttcc
     4801 ggttggtctt gaacagcagg tccacgatgg cctttttctg ctcgccgctc aggaaggcgg
     4861 gctttctcat tccctcggtc acgtatttca ctttggtcag ctcgttatac acggtgaagt
     4921 actcgtacag caggctgtgc ttgggcagca ccttctcgtt gggcaggttc ttatcgaagt
     4981 tggtcatccg ctcgatgaag ctctgggcgg aagcgccctt gtccaccact tcctcgaagt
     5041 tccagggggt gatggtttcc tcgctctttc tggtcatcca ggcgaatctg ctgtttcccc
     5101 tggccagagg gcccacgtag taggggatgc ggaaggtcag gatcttctcg atcttttccc
     5161 ggttgtcctt caggaatggg taaaaatctt cctgccgccg cagaatggcg tgcagctctc
     5221 ccaggtggat ctggtggggg atgctgccgt tgtcgaaggt ccgctgcttc cgcagcaggt
     5281 cctctctgtt cagcttcacg agcagttcct cggtgccgtc catcttttcc aggatgggct
     5341 tgatgaactt gtagaactct tcctggctgg ctccgccgtc aatgtagccg gcgtagccgt
     5401 tcttgctctg gtcgaagaaa atctctttgt acttctcagg cagctgctgc cgcacgagag
     5461 ctttcagcag ggtcaggtcc tggtggtgct cgtcgtatct cttgatcata gaggcgctca
     5521 ggggggcctt ggtgatctcg gtgttcactc tcaggatgtc gctcagcagg atggcgtcgg
     5581 acaggttctt ggcggccaga aacaggtcgg cgtactggtc gccgatctgg gccagcaggt
     5641 tgtccaggtc gtcgtcgtag gtgtccttgc tcagctgcag tttggcatcc tcggccaggt
     5701 cgaagttgct cttgaagttg ggggtcaggc ccaggctcag ggcaatcagg tttccgaaca
     5761 ggccattctt cttctcgccg ggcagctggg cgatcagatt ttccagccgt ctgctcttgc
     5821 tcagtctggc agacaggatg gccttggcgt ccacgccgct ggcgttgatg gggttttcct
     5881 cgaacagctg gttgtaggtc tgcaccagct ggatgaacag cttgtccacg tcgctgttgt
     5941 cggggttcag gtcgccctcg atcaggaagt ggccccggaa cttgatcatg tgggccaggg
     6001 ccagatagat cagccgcagg tcggccttgt cggtgctgtc caccagtttc tttctcaggt
     6061 ggtagatggt ggggtacttc tcgtggtagg ccacctcgtc cacgatgttg ccgaagatgg
     6121 ggtgccgctc gtgcttctta tcctcttcca ccaggaagga ctcttccagt ctgtggaaga
     6181 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt
     6241 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt
     6301 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt
     6361 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc
     6421 ccacagagtt ggtgccgatg tccaggccga tgctgtactt cttgtcgact ttccgcttct
     6481 tctttggtga ctcgaactcg cttccgtcgg ctgtccgttt catatgtata tctccttctt
     6541 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
     6601 gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc ctctacgccg gacgcatcgt
     6661 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     6721 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     6781 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc
     6841 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     6901 taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg
     6961 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     7021 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     7081 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     7141 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     7201 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     7261 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     7321 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     7381 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     7441 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     7501 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     7561 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     7621 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     7681 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     7741 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     7801 acatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     7861 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     7921 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     7981 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8041 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8101 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     8161 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     8221 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     8281 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     8341 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     8401 gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     8461 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     8521 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     8581 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     8641 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     8701 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     8761 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     8821 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     8881 acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     8941 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     9001 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc
     9061 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     9121 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     9181 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     9241 cggcatcaga gcagattgta ctgagagtgc accattgcgg tgtgaaatac cgcacagatg
     9301 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     9361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     9421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     9481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     9541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     9601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     9661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     9721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     9781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     9841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     9901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     9961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    10021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    10081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    10141 gaacgaaaac tcacgttaag ggattttggt catgaacaat aaaactgtct gcttacataa
    10201 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tctaggccgc
    10261 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
    10321 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
    10381 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
    10441 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
    10501 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
    10561 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
    10621 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
    10681 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
    10741 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
    10801 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
    10861 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
    10921 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
    10981 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaagaat
    11041 taattcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    11101 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
    11161 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
    11221 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
    11281 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
    11341 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
    11401 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
    11461 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
    11521 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
    11581 cgtcccattc gcca
//
