LOCUS       sequence-298901-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized Sniper-SpCas9 and blasticidin 
            resistance: EFS promoter-Sniper-Cas9-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12838)
  AUTHORS   Kim N, Kim HK, Lee S, Seo JH, Choi JW, Park J, Min S, Yoon S, Cho 
            SR, Kim HH
  TITLE     Prediction of the sequence-specific cleavage activity of Cas9 
            variants.
  JOURNAL   Nat Biotechnol. 2020 Nov;38(11):1328-1336. doi: 
            10.1038/s41587-020-0537-9. Epub 2020 Jun 8.
  PUBMED    32514125
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: 
            "10.1038/s41587-020-0537-9"; journalName: "Nat Biotechnol"; date: 
            "2020-11"; volume: "38"; issue: "11"; pages: "1328-1336"
COMMENT     Sequence Label: Sniper-Cas9
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    223..340
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        433..644
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      663..672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             669..4772
                     /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
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPASLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEIARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNANLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4773..4820
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4821..4844
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4854..4910
                     /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             4911..5309
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5334..5922
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5387..5407)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5805..5816)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5925..5941)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5926..5942)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6447..6627
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6653..6670)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6659..6883
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6929..7357
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7016..7035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7226..7247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7366..7386)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7531..7665
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7592..7611
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7727..7774
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7793..8167
                     /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    8297..8418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8334..8353)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8388..8407
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8467..8483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8467..8483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8480..8502)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8491..8507
                     /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(8515..8521,8522..8539,8540..8545))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8560..8581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8698..8715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8869..9457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8949..8968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9628..10419,10420..10488))
                     /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     10251..10270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10489..10593)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10668..10687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10859..11238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11240..11438
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11393..11413
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11456..11636
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11683..11808
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12301..12534
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12719..12763
                     /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"
ORIGIN
        1 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
       61 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
      121 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
      181 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
      241 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      301 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
      361 cagagatcca gtttggttaa ttagctagct aggtcttgaa aggagtggga attggctccg
      421 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
      481 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      541 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      601 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttctagagcg
      661 ctgccaccat ggacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
      721 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
      781 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
      841 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
      901 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
      961 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     1021 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     1081 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     1141 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     1201 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     1261 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     1321 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     1381 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     1441 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     1501 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     1561 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     1621 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     1681 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     1741 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     1801 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     1861 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     1921 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     1981 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     2041 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     2101 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     2161 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     2221 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     2281 ccagcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     2341 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     2401 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     2461 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     2521 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     2581 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     2641 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     2701 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     2761 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     2821 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     2881 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     2941 acatcgtgat cgaaatcgcc agagagaacc agaccaccca gaagggacag aagaacagcc
     3001 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     3061 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     3121 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     3181 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     3241 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     3301 agatgaagaa ctactggcgg cagctgctga acgccaacct gattacccag agaaagttcg
     3361 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     3421 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     3481 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     3541 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     3601 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     3661 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     3721 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     3781 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     3841 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     3901 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     3961 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     4021 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     4081 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     4141 agagtgtgaa agagctactg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     4201 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     4261 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     4321 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     4381 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     4441 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     4501 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     4561 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     4621 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     4681 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     4741 cacggatcga cctgtctcag ctgggaggcg acaagcgacc tgccgccaca aagaaggctg
     4801 gacaggctaa gaagaagaaa gattacaaag acgatgacga taagggatcc ggcgcaacaa
     4861 acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atggccaagc
     4921 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     4981 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     5041 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     5101 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     5161 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     5221 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     5281 tgccctctgg ttatgtgtgg gagggctaag aattcgatat caagcttatc ggtaatcaac
     5341 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5401 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5461 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5521 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5581 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5641 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5701 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5761 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5821 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5881 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     5941 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     6001 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     6061 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6121 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     6181 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     6241 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     6301 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     6361 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     6421 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6481 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6541 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6601 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     6661 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     6721 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     6781 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     6841 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     6901 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     6961 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     7021 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     7081 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     7141 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     7201 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     7261 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     7321 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     7381 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     7441 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     7501 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc agttccgccc
     7561 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     7621 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     7681 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     7741 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     7801 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     7861 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     7921 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     7981 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     8041 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     8101 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     8161 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     8221 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     8281 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8341 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8401 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     8461 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     8521 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     8581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     8701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     9001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9061 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     9241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     9301 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     9361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     9421 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     9481 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     9541 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9601 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     9661 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9721 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9781 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9841 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9901 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9961 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    10021 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    10081 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10141 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10201 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    10261 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    10321 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    10381 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    10441 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    10501 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    10561 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    10621 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
    10681 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    10741 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    10801 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
    10861 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
    10921 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
    10981 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
    11041 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
    11101 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
    11161 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
    11221 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
    11281 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
    11341 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
    11401 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
    11461 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    11521 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    11581 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
    11641 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
    11701 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
    11761 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
    11821 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
    11881 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
    11941 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
    12001 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
    12061 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
    12121 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
    12181 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
    12241 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
    12301 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
    12361 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
    12421 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
    12481 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
    12541 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
    12601 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
    12661 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
    12721 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
    12781 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggt
//
