LOCUS       sequence-317692-       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"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2413..2624
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2680..2700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2701..5214,5215..5217,5218..6801)
                     /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             2701..6801
                     /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             8974..8994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9004..9060
                     /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             9061..9459
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9484..10072
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9537..9557)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9955..9966)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10075..10091)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10076..10092)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10597..10777
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10803..10820)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10809..11033
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11079..11507
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11166..11185)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11376..11397
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11516..11536)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11681..11815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11742..11761
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11877..11924
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11943..12317
                     /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    12447..12568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12484..12503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12538..12557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12617..12633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12617..12633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12630..12652)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12641..12657
                     /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(12665..12671,12672..12689,12690..12695))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12710..12731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12848..12865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13019..13607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13099..13118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13778..14569,14570..14638))
                     /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     14401..14420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14639..14743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14818..14837)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttagctagct aggtcttgaa aggagtggga attggctccg
     2401 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     2461 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     2521 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     2581 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttctagagcc
     2641 accatgaaac ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc
     2701 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     2761 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     2821 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     2881 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     2941 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     3001 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     3061 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     3121 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     3181 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     3241 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3301 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3361 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     3421 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     3481 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3541 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3601 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3661 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     3721 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     3781 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     3841 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     3901 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     3961 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     4021 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     4081 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     4141 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     4201 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     4261 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4321 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4381 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4441 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     4501 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4561 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4621 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4681 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     4741 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     4801 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     4861 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     4921 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     4981 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     5041 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     5101 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     5161 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggacgctatc
     5221 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     5281 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     5341 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5401 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     5461 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     5521 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5581 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5641 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     5701 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     5761 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     5821 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     5881 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     5941 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     6001 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     6061 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     6121 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     6181 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     6241 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6301 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     6361 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6421 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     6481 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6541 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6601 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6661 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     6721 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     6781 ctgtctcagc tgggaggtga ctctggagga tctagcggag gatcctctgg cagcgagaca
     6841 ccaggaacaa gcgagtcagc aacaccagag agcagtggcg gcagcagcgg cggcagcagc
     6901 accctaaata tagaagatga gtatcggcta catgagacct caaaagagcc agatgtttct
     6961 ctagggtcca catggctgtc tgattttcct caggcctggg cggaaaccgg gggcatggga
     7021 ctggcagttc gccaagctcc tctgatcata cctctgaaag caacctctac ccccgtgtcc
     7081 ataaaacaat accccatgtc acaagaagcc agactgggga tcaagcccca catacagaga
     7141 ctgttggacc agggaatact ggtaccctgc cagtccccct ggaacacgcc cctgctaccc
     7201 gttaagaaac cagggactaa tgattatagg cctgtccagg atctgagaga agtcaacaag
     7261 cgggtggaag acatccaccc caccgtgccc aacccttaca acctcttgag cgggctccca
     7321 ccgtcccacc agtggtacac tgtgcttgat ttaaaggatg cctttttctg cctgagactc
     7381 caccccacca gtcagcctct cttcgccttt gagtggagag atccagagat gggaatctca
     7441 ggacaattga cctggaccag actcccacag ggtttcaaaa acagtcccac cctgtttaat
     7501 gaggcactgc acagagacct agcagacttc cggatccagc acccagactt gatcctgcta
     7561 cagtacgtgg atgacttact gctggccgcc acttctgagc tagactgcca acaaggtact
     7621 cgggccctgt tacaaaccct agggaacctc gggtatcggg cctcggccaa gaaagcccaa
     7681 atttgccaga aacaggtcaa gtatctgggg tatcttctaa aagagggtca gagatggctg
     7741 actgaggcca gaaaagagac tgtgatgggg cagcctactc cgaagacccc tcgacaacta
     7801 agggagttcc tagggaaggc aggcttctgt cgcctcttca tccctgggtt tgcagaaatg
     7861 gcagcccccc tgtaccctct caccaaaccg gggactctgt ttaattgggg cccagaccaa
     7921 caaaaggcct atcaagaaat caagcaagct cttctaactg ccccagccct ggggttgcca
     7981 gatttgacta agccctttga actctttgtc gacgagaagc agggctacgc caaaggtgtc
     8041 ctaacgcaaa aactgggacc ttggcgtcgg ccggtggcct acctgtccaa aaagctagac
     8101 ccagtagcag ctgggtggcc cccttgccta cggatggtag cagccattgc cgtactgaca
     8161 aaggatgcag gcaagctaac catgggacag ccactagtca ttctggcccc ccatgcagta
     8221 gaggcactag tcaaacaacc ccccgaccgc tggctttcca acgcccggat gactcactat
     8281 caggccttgc ttttggacac ggaccgggtc cagttcggac cggtggtagc cctgaacccg
     8341 gctacgctgc tcccactgcc tgaggaaggg ctgcaacaca actgccttga tatcctggcc
     8401 gaagcccacg gaacccgacc cgacctaacg gaccagccgc tcccagacgc cgaccacacc
     8461 tggtacacgg atggaagcag tctcttacaa gagggacagc gtaaggcggg agctgcggtg
     8521 accaccgaga ccgaggtaat ctgggctaaa gccctgccag ccgggacatc cgctcagcgg
     8581 gctgaactga tagcactcac ccaggcccta aagatggcag aaggtaagaa gctaaatgtt
     8641 tatactgata gccgttatgc ttttgctact gcccatatcc atggagaaat atacagaagg
     8701 cgtgggtggc tcacatcaga aggcaaagag atcaaaaata aagacgagat cttggcccta
     8761 ctaaaagccc tctttctgcc caaaagactt agcataatcc attgtccagg acatcaaaag
     8821 ggacacagcg ccgaggctag aggcaaccgg atggctgacc aagcggcccg aaaggcagcc
     8881 atcacagaga ctccagacac ctctaccctc ctcatagaaa attcatcacc ctctggcggc
     8941 tcaaaaagaa ccgccgacgg cagcgaattc gagcccaaga agaagaggaa agtcggatcc
     9001 ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
     9061 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     9121 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     9181 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     9241 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     9301 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     9361 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     9421 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag aattcgatat caagcttatc
     9481 ggtaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     9541 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     9601 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     9661 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     9721 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     9781 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     9841 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     9901 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     9961 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
    10021 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
    10081 cgtcgacctc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
    10141 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
    10201 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
    10261 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
    10321 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
    10381 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
    10441 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
    10501 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
    10561 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
    10621 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
    10681 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
    10741 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
    10801 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
    10861 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
    10921 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
    10981 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
    11041 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
    11101 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
    11161 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
    11221 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
    11281 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
    11341 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
    11401 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
    11461 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
    11521 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    11581 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
    11641 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    11701 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    11761 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    11821 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
    11881 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
    11941 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    12001 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    12061 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    12121 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    12181 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    12241 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    12301 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
    12361 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
    12421 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
    12481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    12541 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
    12601 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    12661 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    12721 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    12781 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    12841 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    12901 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    12961 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    13021 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    13081 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    13141 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    13201 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    13261 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    13321 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    13381 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    13441 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    13501 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    13561 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    13621 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    13681 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    13741 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    13801 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    13861 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    13921 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    13981 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    14041 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    14101 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    14161 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    14221 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    14281 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    14341 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    14401 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    14461 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    14521 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    14581 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    14641 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    14701 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    14761 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
    14821 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
    14881 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
    14941 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
    15001 tacgcgtt
//
