LOCUS       sequence-458383-       13520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SpG-Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13520)
  AUTHORS   Kaplan EG, Steger RJ, Shah ST, Drepanos LM, Griffith AL, Reint G, 
            Doench JG
  TITLE     Activity-based selection for enhanced base editor mutational 
            scanning.
  JOURNAL   Nat Genet. 2025 Nov;57(11):2920-2929. doi: 
            10.1038/s41588-025-02366-0. Epub 2025 Oct 14.
  PUBMED    41087678
REFERENCE   2  (bases 1 to 13520)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41588-025-02366-0"; journalName: "Nat Genet"; date: 
            "2025-11"; volume: "57"; issue: "11"; pages: "2920-2929"
COMMENT     Sequence Label: pRDB_268
FEATURES             Location/Qualifiers
     source          1..13520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..596
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(61..81)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(479..490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             668..901
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(927..944)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    933..1157
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1203..1631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1290..1309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1500..1521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1640..1660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1645..1974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1825..1960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1887..1906
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2022..2069
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2088..2462
                     /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    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2629..2648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2683..2702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2762..2778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2762..2778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2775..2797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2786..2802
                     /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(2810..2816,2817..2834,2835..2840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2855..2876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2993..3010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3164..3752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3244..3263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3923..4714,4715..4783))
                     /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     4546..4565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4784..4888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4963..4982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5154..5533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5535..5733
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5688..5708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5751..5931
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5978..6103
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6596..6829
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7014..7058
                     /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    7356..7473
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7556..8734
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7787..8725
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8682..8702
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      8739..8748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8784..8804
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8811..12911
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             12918..12965
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12972..13052
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             13059..13112
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             13119..13517
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 attcgaaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
       61 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
      121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
      181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
      241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
      301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
      361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
      421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
      481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
      541 gcgtcctcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
      601 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
      661 gggggactgg aagggctaat tcactcccaa cgaagataag atctgctttt tgcttgtact
      721 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
      781 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      841 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      901 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
      961 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1021 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1081 tggggtgggg caggacagca agggggagga ttgggaagat aatagcaggc atgctgggga
     1141 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     1201 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1261 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1321 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1381 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1441 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1501 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1561 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1621 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1741 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1801 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1861 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     1921 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     1981 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     2041 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     2101 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     2161 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     2221 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     2281 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     2341 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     2401 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     2461 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     2521 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     2581 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2641 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2701 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     2761 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2821 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2881 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2941 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3001 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3061 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3121 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3181 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3241 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3301 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3361 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3421 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3481 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3541 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3601 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3661 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3721 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3781 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3841 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3901 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3961 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4021 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4081 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4141 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4201 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4261 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4321 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4381 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4441 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4501 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4561 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4621 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4681 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4741 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4801 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4861 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     4921 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     4981 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     5041 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     5101 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     5161 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5221 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5281 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     5341 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     5401 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     5461 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     5521 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     5581 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     5641 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     5701 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
     5761 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5821 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5881 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     5941 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     6001 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     6061 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     6121 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     6181 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     6241 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     6301 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     6361 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     6421 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     6481 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     6541 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     6601 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     6661 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     6721 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     6781 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     6841 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     6901 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     6961 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     7021 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     7081 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     7141 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     7201 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     7261 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     7321 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     7381 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     7441 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     7501 ccagtttggt taattaaggt accgggcaga ggtctcgctt ttttcgagac aacacggctc
     7561 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     7621 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     7681 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     7741 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
     7801 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
     7861 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     7921 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     7981 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     8041 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     8101 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     8161 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     8221 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     8281 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     8341 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     8401 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     8461 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     8521 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     8581 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     8641 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     8701 tcaaagtttt tttcttccat ttcaggtgtc gtgaaggagc caccatggcc aaacggacag
     8761 ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcggcagc gacaagaagt
     8821 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     8881 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     8941 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     9001 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     9061 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     9121 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     9181 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     9241 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     9301 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     9361 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgctagcg
     9421 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     9481 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga
     9541 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     9601 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     9661 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     9721 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     9781 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     9841 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     9901 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     9961 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
    10021 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
    10081 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
    10141 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
    10201 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
    10261 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
    10321 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
    10381 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
    10441 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
    10501 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
    10561 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
    10621 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
    10681 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
    10741 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
    10801 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
    10861 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
    10921 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
    10981 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
    11041 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
    11101 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
    11161 aagagggcat caaagagctg ggcagccaga tcctgaaaga acacccagtg gaaaacaccc
    11221 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
    11281 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
    11341 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
    11401 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
    11461 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
    11521 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
    11581 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
    11641 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
    11701 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
    11761 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
    11821 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
    11881 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
    11941 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
    12001 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
    12061 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
    12121 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
    12181 actgggaccc taagaagtac ggcggcttcc tgtggcccac cgtggcctat tctgtgctgg
    12241 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
    12301 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
    12361 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
    12421 agctggaaaa cggccggaag agaatgctgg cctctgccaa gcagctgcag aagggaaacg
    12481 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
    12541 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
    12601 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
    12661 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
    12721 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
    12781 agtactttga caccaccatc gaccggaagc agtacagaag caccaaagag gtgctggacg
    12841 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acgaatcgat ttgtctcagc
    12901 tgggaggcga tgggtccaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
    12961 aaaagggatc ctacccatac gatgttccag attacgctta tccctacgac gtgcctgatt
    13021 atgcataccc atatgatgtc cccgactatg ccggaagcga gggcagagga agccttctaa
    13081 catgcggtga cgtggaggag aatcccggcc cttccgggat ggccaagcct ttgtctcaag
    13141 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgagg
    13201 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
    13261 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
    13321 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
    13381 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
    13441 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
    13501 atgtgtggga gggctaataa
//
