LOCUS       sequence-453897-       11270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes iPE-N prime editor (with MMLV-RT(H8Y, D200N, T306K, W313F, 
            T330P, L603W) fused to the N terminus of nCas9-H840A) driven by a 
            CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11270)
  AUTHORS   Truong DJ, Geilenkeuser J, Wendel SV, Wilming JCH, Armbrust N, 
            Binder EMH, Santl TH, Siebenhaar A, Gruber C, Phlairaharn T, Zivanic
            M, Westmeyer GG
  TITLE     Exonuclease-enhanced prime editors.
  JOURNAL   Nat Methods. 2024 Feb 1. doi: 10.1038/s41592-023-02162-w.
  PUBMED    38302659
REFERENCE   2  (bases 1 to 11270)
  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: "Nat 
            Methods. 2024 Feb 1. doi: 10.1038/s41592-023-02162-w."
COMMENT     Sequence Label: pCAG_iPE-N
FEATURES             Location/Qualifiers
     source          1..11270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     18..34
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        54..433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..714
                     /label=chicken beta-actin promoter
     intron          715..1731
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1655..1677
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1744..1763
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1744..1762
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3819..3839
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3951..6464,6465..6467,6468..8051)
                     /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             3951..8051
                     /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             8064..8084
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8100..8126
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             8139..8165
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             8163..8186
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8207..8223)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8237..8254)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8243..8467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8539..8556)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8539..8555)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8548..8570)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(8696..9151)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8833..8854)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9045..9064
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        9178..9282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9283..9351,9352..10143)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9501..9520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10314..10902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10803..10822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11056..11073
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11190..11211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11226..11231,11232..11249,11250..11256)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(11264..11270,1..10)
                     /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     join(11269..11270,1..21)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 cggataacaa tttcacacag gaaacagcta tgaccatgag gcgcgccgga ttcgacattg
       61 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      121 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      181 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      241 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      301 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      361 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      421 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      481 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
      541 cggggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      601 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      661 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      721 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      781 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      841 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
      901 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
      961 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
     1021 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     1081 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1141 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1201 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1261 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1321 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1381 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1441 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1501 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1561 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1621 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1681 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gatccttaat
     1741 taataatacg actcactata ggggccgcca ccatgaccct gaacatcgag gacgagtacc
     1801 ggctgcacga gacaagcaaa gaacccgatg tgtccctggg cagcacctgg cttagtgatt
     1861 tccctcaggc ctgggccgag acaggcggaa tgggacttgc tgttagacag gcccctctga
     1921 tcatccctct gaaggccaca agcacccctg tgtccatcaa gcagtacccc atgagccaag
     1981 aggcccggct gggaatcaag ccccacattc agagactgct ggaccagggc atcctggtgc
     2041 cttgtcagag cccttggaat acccctctgc tgcccgtgaa gaagcccggc accaacgatt
     2101 acagacccgt gcaggacctg cgggaagtga acaagagagt ggaagatatt caccccaccg
     2161 tgccgaatcc ttacaacctg ctgtctggcc tgcctcctag ccaccagtgg tacacagtgc
     2221 tggacctgaa ggacgccttc ttctgtctgc ggctgcaccc tacaagccag cctctgtttg
     2281 cctttgagtg gcgggaccct gagatgggca ttagcggaca gctgacctgg accagactgc
     2341 cccagggctt caagaacagc cccacactgt tcaacgaggc cctgcatagg gacctcgccg
     2401 acttcagaat ccagcatcct gacctgatcc tgctccagta cgtggacgat ctgctgctgg
     2461 ccgctacaag cgagctggat tgtcagcagg gaacaagagc cctgctgcaa accctgggca
     2521 acctgggcta tagagcctct gccaagaagg cccagatttg ccagaaacaa gtgaagtatc
     2581 tgggctacct gctgaaagag ggccagcgtt ggctgaccga ggccagaaaa gaaaccgtga
     2641 tgggccagcc tacacctaag acacccagac agctgagaga gttcctgggc aaagccggat
     2701 tctgtcggct gttcatccct ggctttgccg agatggctgc ccctctgtac ccactgacaa
     2761 agcccggaac tctgttcaac tggggcccag atcagcagaa ggcctaccaa gagatcaagc
     2821 aggctctgct gacagcccct gctctgggac tgcctgatct gaccaagcct ttcgagctgt
     2881 tcgtggacga gaagcagggc tatgccaaag gcgtgctgac acagaagctc ggcccttgga
     2941 gaaggcctgt ggcctacctg agcaagaaac tggaccctgt ggctgccgga tggcctcctt
     3001 gtctgagaat ggtggccgcc attgccgtgc tgaccaagga tgccggaaag ctgacaatgg
     3061 gacagcctct ggtcattctg gcccctcatg ccgtggaagc cctcgtgaaa cagcctcctg
     3121 atcggtggct gagcaacgcc aggatgacac actatcaggc actgctgctc gacaccgaca
     3181 gagtgcagtt tggacctgtg gtggccctga atcctgccac acttctgcct ctgcctgagg
     3241 aaggcctcca gcacaattgc ctggacatcc tggccgaggc tcacggcaca agacccgatc
     3301 tgacagatca gccactgcct gacgccgacc acacctggta tacagatggc agctctctgc
     3361 tgcaagaagg acagagaaaa gccggggctg ccgtgaccac cgagacagaa gtgatttggg
     3421 ccaaagctct gcccgctggc acatctgctc agagagccga actgatcgcc ctgacacagg
     3481 ccctgaaaat ggccgagggc aagaagctga acgtctacac cgactccaga tacgccttcg
     3541 ccaccgctca catccacggc gaaatctatc ggcggagagg atggctgacc agcgagggca
     3601 aagagattaa gaacaaggac gagattctcg ccctgctcaa ggccctgttc ctgcctaagc
     3661 ggctgagcat catccactgt ccaggccacc agaagggcca ctctgctgaa gctagaggca
     3721 acagaatggc cgaccaggct gccagaaagg ccgccatcac agagacaccc gataccagca
     3781 cactgctgat cgagaacagc agccctggat ccacgccgcc caagaagaag agaaaggttg
     3841 aggacggcga gggcaccggt ggacctggaa gcgagcctgc tacatctggc tctgagacac
     3901 ctggcacctc cgagtctgct acacctgaat ctggacctgg cggatccgga gacaagaagt
     3961 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     4021 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     4081 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     4141 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     4201 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     4261 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     4321 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     4381 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     4441 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     4501 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     4561 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     4621 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga
     4681 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     4741 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     4801 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     4861 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     4921 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     4981 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     5041 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     5101 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     5161 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     5221 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     5281 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     5341 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     5401 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     5461 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     5521 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     5581 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     5641 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     5701 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     5761 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     5821 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     5881 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     5941 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     6001 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     6061 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     6121 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     6181 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     6241 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     6301 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     6361 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     6421 accaggaact ggacatcaac cggctgtccg actacgatgt ggacgccatc gtgcctcaga
     6481 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     6541 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     6601 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     6661 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     6721 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     6781 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     6841 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     6901 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     6961 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     7021 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     7081 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     7141 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     7201 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     7261 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     7321 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     7381 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     7441 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     7501 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     7561 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     7621 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     7681 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     7741 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     7801 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     7861 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     7921 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     7981 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     8041 tgggaggcga cggatccaca ccacctaaga agaaacggaa ggtcgaggac ggcgagggcc
     8101 ctgctgctaa gagagtgaaa ctggactccg gagctgctcc agccgccaag aagaagaagc
     8161 tcgactacaa ggacgacgac gataagtgaa cgcgtaaatg attgcagatc cactagttct
     8221 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     8281 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     8341 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     8401 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     8461 tctatggctt ctgaggcgga aagaaccagc tggggctcga gatccactag ttctagcctc
     8521 gaggctagag cggccgccac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     8581 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     8641 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt
     8701 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     8761 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     8821 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     8881 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     8941 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     9001 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     9061 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     9121 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     9181 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     9241 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     9301 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     9361 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     9421 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     9481 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     9541 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     9601 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     9661 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     9721 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     9781 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     9841 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     9901 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     9961 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
    10021 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    10081 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    10141 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    10201 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    10261 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    10321 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    10381 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    10441 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    10501 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    10561 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    10621 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    10681 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    10741 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    10801 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    10861 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    10921 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    10981 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    11041 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    11101 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    11161 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    11221 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
//
