LOCUS       sequence-395414-       11939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express His-tagged PE2 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11939)
  AUTHORS   Liang SQ, Liu P, Ponnienselvan K, Suresh S, Chen Z, Kramme C, 
            Chatterjee P, Zhu LJ, Sontheimer EJ, Xue W, Wolfe SA
  TITLE     Genome-wide profiling of prime editor off-target sites in vitro and 
            in vivo using PE-tag.
  JOURNAL   Nat Methods. 2023 Jun;20(6):898-907. doi: 
            10.1038/s41592-023-01859-2. Epub 2023 May 8.
  PUBMED    37156841
REFERENCE   2  (bases 1 to 11939)
  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"; doi: 
            "10.1038/s41592-023-01859-2"; journalName: "Nat Methods"; date: 
            "2023-06"; volume: "20"; issue: "6"; pages: "898-907"
COMMENT     Sequence Label: pET-21a-PE2-His
FEATURES             Location/Qualifiers
     source          1..11939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     660..679
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(699..776)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     982..1001
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1085..1104
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1085..1103
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1104..1128
                     /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             1143..1165
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1173..1205
                     /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             1221..1247
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1284..1310
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             join(1362..3875,3876..3878,3879..5462)
                     /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             1362..5462
                     /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             7653..7679
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7722..7748
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7755..7772
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7825..7843)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7839..7886
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      7923..8378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8010..8029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8220..8241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8404..8508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8509..8577,8578..9369)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8727..8746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9540..10128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10029..10048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10282..10299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10314..10453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10379..10398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10539..10561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(10555..10746)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(11555..11939,1..698))
                     /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"
ORIGIN
        1 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
       61 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
      121 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
      181 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
      241 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
      301 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
      361 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
      421 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
      481 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
      541 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
      601 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
      661 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
      721 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
      781 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
      841 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
      901 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
      961 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     1021 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     1081 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     1141 attttgttta actttaagaa ggagatatac atatggctag catgactggt ggacagcaaa
     1201 tgggtcgcgg atccgaattc cccgccgcca agcgcgtgaa gctggacagc ggcagcggcg
     1261 gcggcagcgg cggcggcagc ggcccggcag ctaagagggt gaaactggat ggcggtagcg
     1321 gcggaggcag cggtggcggc agcggccccc tcgaggtcga cgacaagaag tacagcatcg
     1381 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     1441 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     1501 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
     1561 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     1621 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     1681 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     1741 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     1801 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     1861 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     1921 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     1981 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     2041 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
     2101 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     2161 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     2221 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     2281 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     2341 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     2401 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     2461 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     2521 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     2581 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     2641 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     2701 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     2761 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     2821 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     2881 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     2941 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     3001 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     3061 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     3121 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     3181 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     3241 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     3301 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     3361 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     3421 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     3481 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     3541 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     3601 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     3661 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     3721 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     3781 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     3841 tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag agctttctga
     3901 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     3961 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     4021 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     4081 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     4141 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     4201 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     4261 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     4321 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     4381 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     4441 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     4501 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     4561 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
     4621 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     4681 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
     4741 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
     4801 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     4861 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     4921 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     4981 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
     5041 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     5101 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
     5161 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     5221 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
     5281 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     5341 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     5401 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggtg
     5461 actctggagg atctagcgga ggatcctctg gcagcgagac accaggaaca agcgagtcag
     5521 caacaccaga gagcagtggc ggcagcagcg gcggcagcag caccctaaat atagaagatg
     5581 agtatcggct acatgagacc tcaaaagagc cagatgtttc tctagggtcc acatggctgt
     5641 ctgattttcc tcaggcctgg gcggaaaccg ggggcatggg actggcagtt cgccaagctc
     5701 ctctgatcat acctctgaaa gcaacctcta cccccgtgtc cataaaacaa taccccatgt
     5761 cacaagaagc cagactgggg atcaagcccc acatacagag actgttggac cagggaatac
     5821 tggtaccctg ccagtccccc tggaacacgc ccctgctacc cgttaagaaa ccagggacta
     5881 atgattatag gcctgtccag gatctgagag aagtcaacaa gcgggtggaa gacatccacc
     5941 ccaccgtgcc caacccttac aacctcttga gcgggctccc accgtcccac cagtggtaca
     6001 ctgtgcttga tttaaaggat gcctttttct gcctgagact ccaccccacc agtcagcctc
     6061 tcttcgcctt tgagtggaga gatccagaga tgggaatctc aggacaattg acctggacca
     6121 gactcccaca gggtttcaaa aacagtccca ccctgtttaa tgaggcactg cacagagacc
     6181 tagcagactt ccggatccag cacccagact tgatcctgct acagtacgtg gatgacttac
     6241 tgctggccgc cacttctgag ctagactgcc aacaaggtac tcgggccctg ttacaaaccc
     6301 tagggaacct cgggtatcgg gcctcggcca agaaagccca aatttgccag aaacaggtca
     6361 agtatctggg gtatcttcta aaagagggtc agagatggct gactgaggcc agaaaagaga
     6421 ctgtgatggg gcagcctact ccgaagaccc ctcgacaact aagggagttc ctagggaagg
     6481 caggcttctg tcgcctcttc atccctgggt ttgcagaaat ggcagccccc ctgtaccctc
     6541 tcaccaaacc ggggactctg tttaattggg gcccagacca acaaaaggcc tatcaagaaa
     6601 tcaagcaagc tcttctaact gccccagccc tggggttgcc agatttgact aagccctttg
     6661 aactctttgt cgacgagaag cagggctacg ccaaaggtgt cctaacgcaa aaactgggac
     6721 cttggcgtcg gccggtggcc tacctgtcca aaaagctaga cccagtagca gctgggtggc
     6781 ccccttgcct acggatggta gcagccattg ccgtactgac aaaggatgca ggcaagctaa
     6841 ccatgggaca gccactagtc attctggccc cccatgcagt agaggcacta gtcaaacaac
     6901 cccccgaccg ctggctttcc aacgcccgga tgactcacta tcaggccttg cttttggaca
     6961 cggaccgggt ccagttcgga ccggtggtag ccctgaaccc ggctacgctg ctcccactgc
     7021 ctgaggaagg gctgcaacac aactgccttg atatcctggc cgaagcccac ggaacccgac
     7081 ccgacctaac ggaccagccg ctcccagacg ccgaccacac ctggtacacg gatggaagca
     7141 gtctcttaca agagggacag cgtaaggcgg gagctgcggt gaccaccgag accgaggtaa
     7201 tctgggctaa agccctgcca gccgggacat ccgctcagcg ggctgaactg atagcactca
     7261 cccaggccct aaagatggca gaaggtaaga agctaaatgt ttatactgat agccgttatg
     7321 cttttgctac tgcccatatc catggagaaa tatacagaag gcgtgggtgg ctcacatcag
     7381 aaggcaaaga gatcaaaaat aaagacgaga tcttggccct actaaaagcc ctctttctgc
     7441 ccaaaagact tagcataatc cattgtccag gacatcaaaa gggacacagc gccgaggcta
     7501 gaggcaaccg gatggctgac caagcggccc gaaaggcagc catcacagag actccagaca
     7561 cctctaccct cctcatagaa aattcatcac cctctggcgg ctcaaaaaga accgccgacg
     7621 gcagcgaatt cgagtctaga ggatccgccg ctcccgccgc caagcgcgtg aagctggacg
     7681 gcggcagcgg cggcggcagc ggcggcggca gcggctccgg accggcagct aagagggtga
     7741 aactggatct cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     7801 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     7861 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     7921 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     7981 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     8041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8101 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     8161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     8221 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     8281 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     8341 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     8401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     8461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     8521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     8581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     8641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     8701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     8761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     8821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     8881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     8941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     9001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     9061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     9121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     9181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     9241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     9301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     9361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     9421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     9481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     9541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     9601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     9661 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     9721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     9781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     9841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     9901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     9961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
    10021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
    10081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
    10141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
    10201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
    10261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
    10321 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
    10381 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
    10441 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
    10501 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
    10561 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
    10621 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
    10681 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
    10741 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
    10801 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
    10861 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
    10921 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
    10981 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
    11041 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
    11101 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
    11161 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
    11221 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
    11281 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
    11341 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
    11401 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
    11461 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
    11521 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
    11581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    11641 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
    11701 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
    11761 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
    11821 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
    11881 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattc
//
