LOCUS       sequence-388906-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH and blasticidin 
            resistance: EFS promoter-SpCas9-NRCH-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12838)
  AUTHORS   Kim N, Choi S, Kim S, Song M, Seo JH, Min S, Park J, Cho SR, Kim HH
  TITLE     Deep learning models to predict the editing efficiencies and 
            outcomes of diverse base editors.
  JOURNAL   Nat Biotechnol. 2024 Mar;42(3):484-497. doi: 
            10.1038/s41587-023-01792-x
  PUBMED    37188916
REFERENCE   2  (bases 1 to 12838)
  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"; journalName: 
            "Nat Biotechnol"; date: "2024-03"; volume: "42"; issue: "3"; pages: 
            "484-497"
COMMENT     Sequence Label: SpCas9-NRCH
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..691
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     646..666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             709..889
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    936..1061
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1554..1787
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1972..2016
                     /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    2314..2431
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2524..2735
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2754..2763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2760..6863
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGG
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKGNSDKLIARKKDWDPKKYGGFNSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTINRKQYNTTKEVL
                     DATLIRQSITGLYETRIDLSQLGGD"
     CDS             6864..6911
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6912..6935
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6945..7001
                     /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             7002..7400
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7425..8013
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7478..7498)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7896..7907)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8016..8032)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8017..8033)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8538..8718
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8744..8761)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8750..8974
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9020..9448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9107..9126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9317..9338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9457..9477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      9622..9756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9683..9702
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9818..9865
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9884..10258
                     /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    10388..10509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10425..10444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10479..10498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10558..10574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10558..10574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10571..10593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10582..10598
                     /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(10606..10612,10613..10630,10631..10636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10651..10672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10789..10806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10960..11548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11040..11059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11719..12510,12511..12579))
                     /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     12342..12361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12580..12684)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12759..12778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
       61 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      121 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      661 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      721 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      781 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      841 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      901 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      961 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1021 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1081 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1141 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1201 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1261 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1321 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1381 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1441 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1501 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1561 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1621 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1681 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1741 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1801 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1861 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1921 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1981 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     2041 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2101 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     2161 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     2221 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     2281 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     2341 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     2401 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     2461 agtttggtta attagctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     2521 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     2581 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     2641 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     2701 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca
     2761 tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     2821 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     2881 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     2941 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     3001 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     3061 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     3121 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     3181 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     3241 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     3301 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     3361 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     3421 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     3481 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     3541 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     3601 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     3661 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     3721 tggtgaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     3781 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     3841 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     3901 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3961 agcagcggac cttcgacaac ggcatcatcc cccaccagat ccacctggga gagctgcacg
     4021 ccattctgcg gcggcagggc gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     4081 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     4141 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     4201 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     4261 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     4321 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     4381 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     4441 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     4501 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     4561 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     4621 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     4681 acctgttcga cgacaaagtg atgaagcagc tgaagcggct gagatacacc ggctggggca
     4741 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     4801 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     4861 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     4921 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     4981 tgaaggtggt ggacgagctc gtgaaagtga tgggcggcca caagcccgag aacatcgtga
     5041 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     5101 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     5161 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     5221 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     5281 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     5341 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     5401 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     5461 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     5521 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     5581 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     5641 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     5701 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     5761 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     5821 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     5881 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     5941 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     6001 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     6061 agacaggcgg cttcagcaaa gagtctatcc tgcccaaggg caacagcgat aagctgatcg
     6121 ccagaaagaa ggactgggac cctaagaagt acggcggctt caacagcccc accgtggcct
     6181 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     6241 aagagctact ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     6301 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     6361 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgtgctgc
     6421 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     6481 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     6541 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     6601 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     6661 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     6721 ctgccgcctt caagtacttt gacaccacca tcaaccggaa gcagtacaac accaccaaag
     6781 aggtgctgga cgccaccctg atccggcaga gcatcaccgg cctgtacgag acacggatcg
     6841 acctgtctca gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta
     6901 agaagaagaa agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc
     6961 tgctgaaaca agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc
     7021 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     7081 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     7141 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     7201 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     7261 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     7321 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     7381 gttatgtgtg ggagggctaa gaattcgata tcaagcttat cggtaatcaa cctctggatt
     7441 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     7501 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     7561 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     7621 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     7681 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     7741 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     7801 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     7861 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     7921 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     7981 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
     8041 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     8101 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     8161 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     8221 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     8281 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     8341 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     8401 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     8461 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     8521 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8581 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8641 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8701 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     8761 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     8821 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     8881 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     8941 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     9001 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     9061 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     9121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     9181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     9241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     9301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     9361 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     9421 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     9481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     9541 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     9601 tcaattagtc agcaaccata gtcccgcccc taactccgcc cagttccgcc cattctccgc
     9661 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     9721 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     9781 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     9841 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     9901 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     9961 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    10021 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    10081 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    10141 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    10201 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    10261 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    10321 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    10381 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    10441 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    10501 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    10561 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    10621 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    10681 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    10741 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    10801 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    10861 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    10921 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    10981 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    11041 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    11101 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    11161 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    11221 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    11281 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    11341 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    11401 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    11461 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    11521 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    11581 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    11641 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    11701 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    11761 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    11821 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    11881 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    11941 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    12001 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    12061 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    12121 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    12181 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    12241 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    12301 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    12361 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    12421 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    12481 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    12541 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    12601 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    12661 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
    12721 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
    12781 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcga
//
