LOCUS       sequence-388886-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRTH and blasticidin 
            resistance: EFS promoter-SpCas9-NRTH-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-NRTH
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    417..538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(454..473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     508..527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(587..603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(587..603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(600..622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    611..627
                     /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(635..641,642..659,660..665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    680..701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(818..835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(989..1577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1069..1088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1748..2539,2540..2608))
                     /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     2371..2390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2609..2713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2788..2807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2979..3358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3360..3558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3513..3533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3576..3756
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3803..3928
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4421..4654
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4839..4883
                     /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    5181..5298
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5391..5602
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5621..5630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5627..9730
                     /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
                     TIMERSSFEKNPIGFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASASVLHKGNE
                     LALPSKYVNFLYLASHYEKLKGSSEDNKQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGASAAFKYFDTTIGRKLYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9731..9778
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9779..9802
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9812..9868
                     /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             9869..10267
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    10292..10880
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10345..10365)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10763..10774)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10883..10899)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10884..10900)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11405..11585
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11611..11628)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11617..11841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11887..12315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11974..11993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12184..12205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12324..12344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      12489..12623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12550..12569
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12685..12732
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(12751..12838,1..287)
                     /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"
ORIGIN
        1 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
       61 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
      121 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
      181 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
      241 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
      301 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
      361 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
      421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
      481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
      541 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
      601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      661 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     2761 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2821 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     2881 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     2941 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3001 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3061 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     3361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     3421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     3481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     3541 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     3601 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3661 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3721 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     3781 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     3841 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     3901 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     3961 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     4021 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     4081 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     4141 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     4201 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     4261 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     4321 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     4381 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     4441 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     4501 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     4561 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     4621 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     4681 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     4741 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     4801 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     4861 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     4921 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     4981 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     5041 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     5101 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     5161 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     5221 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     5281 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     5341 agctagctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     5401 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta
     5461 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     5521 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     5581 cgggtttgcc gccagaacac aggaccggtt ctagagcgct gccaccatgg acaagaagta
     5641 cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     5701 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     5761 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     5821 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     5881 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     5941 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     6001 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     6061 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     6121 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     6181 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     6241 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     6301 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     6361 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     6421 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     6481 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     6541 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatgg tgaagagata
     6601 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     6661 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     6721 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     6781 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     6841 cgacaacggc atcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     6901 gcagggcgat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     6961 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     7021 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     7081 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     7141 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     7201 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     7261 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     7321 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     7381 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     7441 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     7501 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     7561 caaagtgatg aagcagctga agcggctgag atacaccggc tggggcaggc tgagccggaa
     7621 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     7681 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     7741 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     7801 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     7861 cgagctcgtg aaagtgatgg gcggccacaa gcccgagaac atcgtgatcg aaatggccag
     7921 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     7981 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     8041 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     8101 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     8161 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     8221 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     8281 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     8341 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     8401 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     8461 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     8521 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     8581 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     8641 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     8701 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     8761 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     8821 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     8881 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     8941 cagcaaagag tctatcctgc ccaagggcaa cagcgataag ctgatcgcca gaaagaagga
     9001 ctgggaccct aagaagtacg gcggcttcaa cagccccacc gtggcctatt ctgtgctggt
     9061 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctactggg
     9121 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcggctttc tggaagccaa
     9181 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     9241 gctggaaaac ggccggaaga gaatgctggc ctctgccagc gtgctgcaca agggaaacga
     9301 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     9361 gaagggctcc agcgaggata ataagcagaa acagctgttt gtggaacagc acaagcacta
     9421 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     9481 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     9541 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagccagcg ccgccttcaa
     9601 gtactttgac accaccatcg gccggaagct gtacaccagc accaaagagg tgctggacgc
     9661 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     9721 gggaggcgac aagcgacctg ccgccacaaa gaaggctgga caggctaaga agaagaaaga
     9781 ttacaaagac gatgacgata agggatccgg cgcaacaaac ttctctctgc tgaaacaagc
     9841 cggagatgtc gaagagaatc ctggaccgat ggccaagcct ttgtctcaag aagaatccac
     9901 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     9961 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
    10021 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
    10081 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
    10141 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
    10201 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
    10261 gggctaagaa ttcgatatca agcttatcgg taatcaacct ctggattaca aaatttgtga
    10321 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
    10381 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
    10441 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
    10501 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
    10561 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
    10621 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
    10681 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
    10741 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
    10801 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
    10861 cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc
    10921 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
    10981 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
    11041 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
    11101 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
    11161 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
    11221 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
    11281 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
    11341 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
    11401 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
    11461 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
    11521 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
    11581 tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
    11641 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
    11701 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
    11761 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
    11821 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
    11881 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
    11941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
    12001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
    12061 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
    12121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
    12181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
    12241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
    12301 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
    12361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
    12421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
    12481 aaccatagtc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
    12541 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
    12601 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
    12661 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
    12721 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
    12781 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttct
//
