LOCUS       sequence-312966-       15008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PE2 in mammalian cells. Can be used to generate stable
            cell lines by Blasticidin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15008)
  AUTHORS   Kim HK, Yu G, Park J, Min S, Lee S, Yoon S, Kim HH
  TITLE     Predicting the efficiency of prime editing guide RNAs in human 
            cells.
  JOURNAL   Nat Biotechnol. 2020 Sep 21. pii: 10.1038/s41587-020-0677-y. doi: 
            10.1038/s41587-020-0677-y.
  PUBMED    32958957
REFERENCE   2  (bases 1 to 15008)
  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 
            Biotechnol. 2020 Sep 21. pii: 10.1038/s41587-020-0677-y. doi: 
            10.1038/s41587-020-0677-y."
COMMENT     Sequence Label: pLenti-PE2-BSD
FEATURES             Location/Qualifiers
     source          1..15008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             119..299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(325..342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    331..555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      601..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(688..707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     898..919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1038..1058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1203..1337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1264..1283
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1399..1446
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1465..1839
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1969..2090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2006..2025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2060..2079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2139..2155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2139..2155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2152..2174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2163..2179
                     /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)."
     promoter        complement(join(2187..2193,2194..2211,2212..2217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2232..2253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2621..2640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3300..4091,4092..4160))
                     /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     3923..3942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4161..4265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4340..4359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4531..4910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4912..5110
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5065..5085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5128..5308
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5355..5480
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5973..6206
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6391..6435
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6733..6850
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6943..7154
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7210..7230
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7231..9744,9745..9747,9748..11331)
                     /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             7231..11331
                     /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             13504..13524
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13534..13590
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             13591..13989
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    14014..14602
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14067..14087)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14485..14496)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(14605..14621)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14606..14622)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
       61 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
      121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      301 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
      361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
      421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
      481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
      541 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
      601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      961 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1201 agtcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1261 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     1321 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     1381 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     1441 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     1501 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     1561 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     1621 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     1681 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     1741 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     1801 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     1861 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1921 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1981 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2101 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2161 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2221 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2281 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4321 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4381 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4441 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4501 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4561 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4681 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4741 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4801 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4861 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4921 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4981 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5041 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5101 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     5161 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5221 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5281 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     5341 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     5401 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     5461 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     5521 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     5581 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     5641 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     5701 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     5761 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     5821 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     5881 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     5941 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     6001 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     6061 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     6121 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     6181 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     6241 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     6301 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     6361 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     6421 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     6481 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     6541 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     6601 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     6661 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
     6721 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     6781 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     6841 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttagctagct
     6901 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
     6961 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
     7021 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     7081 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     7141 ccgccagaac acaggaccgg ttctagagcc accatgaaac ggacagccga cggaagcgag
     7201 ttcgagtcac caaagaagaa gcggaaagtc gacaagaagt acagcatcgg cctggacatc
     7261 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     7321 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     7381 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     7441 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     7501 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     7561 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     7621 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     7681 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     7741 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     7801 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     7861 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     7921 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     7981 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     8041 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     8101 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     8161 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     8221 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     8281 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     8341 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     8401 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     8461 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     8521 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     8581 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     8641 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     8701 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     8761 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     8821 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     8881 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     8941 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     9001 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     9061 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     9121 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     9181 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     9241 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     9301 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     9361 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     9421 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     9481 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     9541 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     9601 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     9661 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     9721 cggctgtccg actacgatgt ggacgctatc gtgcctcaga gctttctgaa ggacgactcc
     9781 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     9841 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     9901 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     9961 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
    10021 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
    10081 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
    10141 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
    10201 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
    10261 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
    10321 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
    10381 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
    10441 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
    10501 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
    10561 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
    10621 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
    10681 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
    10741 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
    10801 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
    10861 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
    10921 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
    10981 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
    11041 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
    11101 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
    11161 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
    11221 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
    11281 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggtga ctctggagga
    11341 tctagcggag gatcctctgg cagcgagaca ccaggaacaa gcgagtcagc aacaccagag
    11401 agcagtggcg gcagcagcgg cggcagcagc accctaaata tagaagatga gtatcggcta
    11461 catgagacct caaaagagcc agatgtttct ctagggtcca catggctgtc tgattttcct
    11521 caggcctggg cggaaaccgg gggcatggga ctggcagttc gccaagctcc tctgatcata
    11581 cctctgaaag caacctctac ccccgtgtcc ataaaacaat accccatgtc acaagaagcc
    11641 agactgggga tcaagcccca catacagaga ctgttggacc agggaatact ggtaccctgc
    11701 cagtccccct ggaacacgcc cctgctaccc gttaagaaac cagggactaa tgattatagg
    11761 cctgtccagg atctgagaga agtcaacaag cgggtggaag acatccaccc caccgtgccc
    11821 aacccttaca acctcttgag cgggctccca ccgtcccacc agtggtacac tgtgcttgat
    11881 ttaaaggatg cctttttctg cctgagactc caccccacca gtcagcctct cttcgccttt
    11941 gagtggagag atccagagat gggaatctca ggacaattga cctggaccag actcccacag
    12001 ggtttcaaaa acagtcccac cctgtttaat gaggcactgc acagagacct agcagacttc
    12061 cggatccagc acccagactt gatcctgcta cagtacgtgg atgacttact gctggccgcc
    12121 acttctgagc tagactgcca acaaggtact cgggccctgt tacaaaccct agggaacctc
    12181 gggtatcggg cctcggccaa gaaagcccaa atttgccaga aacaggtcaa gtatctgggg
    12241 tatcttctaa aagagggtca gagatggctg actgaggcca gaaaagagac tgtgatgggg
    12301 cagcctactc cgaagacccc tcgacaacta agggagttcc tagggaaggc aggcttctgt
    12361 cgcctcttca tccctgggtt tgcagaaatg gcagcccccc tgtaccctct caccaaaccg
    12421 gggactctgt ttaattgggg cccagaccaa caaaaggcct atcaagaaat caagcaagct
    12481 cttctaactg ccccagccct ggggttgcca gatttgacta agccctttga actctttgtc
    12541 gacgagaagc agggctacgc caaaggtgtc ctaacgcaaa aactgggacc ttggcgtcgg
    12601 ccggtggcct acctgtccaa aaagctagac ccagtagcag ctgggtggcc cccttgccta
    12661 cggatggtag cagccattgc cgtactgaca aaggatgcag gcaagctaac catgggacag
    12721 ccactagtca ttctggcccc ccatgcagta gaggcactag tcaaacaacc ccccgaccgc
    12781 tggctttcca acgcccggat gactcactat caggccttgc ttttggacac ggaccgggtc
    12841 cagttcggac cggtggtagc cctgaacccg gctacgctgc tcccactgcc tgaggaaggg
    12901 ctgcaacaca actgccttga tatcctggcc gaagcccacg gaacccgacc cgacctaacg
    12961 gaccagccgc tcccagacgc cgaccacacc tggtacacgg atggaagcag tctcttacaa
    13021 gagggacagc gtaaggcggg agctgcggtg accaccgaga ccgaggtaat ctgggctaaa
    13081 gccctgccag ccgggacatc cgctcagcgg gctgaactga tagcactcac ccaggcccta
    13141 aagatggcag aaggtaagaa gctaaatgtt tatactgata gccgttatgc ttttgctact
    13201 gcccatatcc atggagaaat atacagaagg cgtgggtggc tcacatcaga aggcaaagag
    13261 atcaaaaata aagacgagat cttggcccta ctaaaagccc tctttctgcc caaaagactt
    13321 agcataatcc attgtccagg acatcaaaag ggacacagcg ccgaggctag aggcaaccgg
    13381 atggctgacc aagcggcccg aaaggcagcc atcacagaga ctccagacac ctctaccctc
    13441 ctcatagaaa attcatcacc ctctggcggc tcaaaaagaa ccgccgacgg cagcgaattc
    13501 gagcccaaga agaagaggaa agtcggatcc ggcgcaacaa acttctctct gctgaaacaa
    13561 gccggagatg tcgaagagaa tcctggaccg atggccaagc ctttgtctca agaagaatcc
    13621 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
    13681 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
    13741 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
    13801 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
    13861 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
    13921 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
    13981 gagggctaag aattcgatat caagcttatc ggtaatcaac ctctggatta caaaatttgt
    14041 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
    14101 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
    14161 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
    14221 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
    14281 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
    14341 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
    14401 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
    14461 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
    14521 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
    14581 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga
    14641 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
    14701 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
    14761 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
    14821 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
    14881 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
    14941 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
    15001 acccgctt
//
