LOCUS       sequence-274786-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-VRQR and blasticidin 
            resistance: EFS promoter-VRQR-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: VRQR variant
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    193..310
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        403..614
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      633..642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             639..4742
                     /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
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4743..4790
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4791..4814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4824..4880
                     /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             4881..5279
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5304..5892
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5357..5377)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5775..5786)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5895..5911)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5896..5912)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6417..6597
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6623..6640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6629..6853
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6899..7327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6986..7005)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7196..7217
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7336..7356)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7501..7635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7562..7581
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7697..7744
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7763..8137
                     /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    8267..8388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8304..8323)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8358..8377
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8437..8453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8437..8453)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8450..8472)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8461..8477
                     /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(8485..8491,8492..8509,8510..8515))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8530..8551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8668..8685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8839..9427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8919..8938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9598..10389,10390..10458))
                     /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     10221..10240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10459..10563)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10638..10657)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10829..11208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11210..11408
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11363..11383
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11426..11606
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11653..11778
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12271..12504
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12689..12733
                     /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 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
       61 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
      121 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
      181 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
      241 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
      301 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttagctagct
      361 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      421 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
      481 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      541 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      601 ccgccagaac acaggaccgg ttctagagcg ctgccaccat ggacaagaag tacagcatcg
      661 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
      721 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
      781 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
      841 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
      901 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
      961 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     1021 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     1081 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     1141 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     1201 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     1261 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     1321 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
     1381 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     1441 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     1501 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     1561 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     1621 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     1681 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     1741 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     1801 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     1861 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     1921 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     1981 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     2041 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     2101 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     2161 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     2221 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     2281 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     2341 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     2401 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     2461 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     2521 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     2581 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     2641 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     2701 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     2761 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     2821 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     2881 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     2941 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     3001 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     3061 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     3121 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga
     3181 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     3241 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     3301 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     3361 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     3421 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     3481 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     3541 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     3601 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     3661 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     3721 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     3781 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     3841 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
     3901 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     3961 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
     4021 ctaagaagta cggcggcttc gtgagcccca ccgtggccta ttctgtgctg gtggtggcca
     4081 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctactg gggatcacca
     4141 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     4201 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     4261 acggccggaa gagaatgctg gcctctgccc gggaactgca gaagggaaac gaactggccc
     4321 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     4381 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
     4441 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     4501 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
     4561 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     4621 acaccaccat cgaccggaag cagtaccgga gcaccaaaga ggtgctggac gccaccctga
     4681 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
     4741 acaagcgacc tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag
     4801 acgatgacga taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     4861 tcgaagagaa tcctggaccg atggccaagc ctttgtctca agaagaatcc accctcattg
     4921 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     4981 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     5041 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     5101 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     5161 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     5221 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag
     5281 aattcgatat caagcttatc ggtaatcaac ctctggatta caaaatttgt gaaagattga
     5341 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5401 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5461 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5521 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5581 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5641 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     5701 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5761 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5821 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5881 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa
     5941 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     6001 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     6061 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6121 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     6181 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     6241 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     6301 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     6361 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     6421 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6481 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6541 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
     6601 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6661 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6721 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6781 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6841 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     6901 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6961 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7021 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7081 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7141 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7201 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7261 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7321 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     7381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     7441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7501 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     7561 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     7621 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     7681 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     7741 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     7801 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     7861 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     7921 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     7981 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     8041 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     8101 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     8161 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     8221 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     8281 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     8341 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     8401 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     8461 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     8521 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     8581 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     8641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     9001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     9061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     9121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     9181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     9241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     9301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     9361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     9421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     9481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     9541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     9601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     9661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     9721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     9781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
    10021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
    10081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
    10141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
    10201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
    10261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
    10321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
    10381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
    10441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
    10501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
    10561 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
    10621 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
    10681 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
    10741 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
    10801 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
    10861 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
    10921 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
    10981 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
    11041 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
    11101 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
    11161 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
    11221 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
    11281 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
    11341 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
    11401 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
    11461 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
    11521 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
    11581 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
    11641 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
    11701 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
    11761 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
    11821 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
    11881 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
    11941 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
    12001 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
    12061 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
    12121 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
    12181 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
    12241 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
    12301 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
    12361 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
    12421 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
    12481 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
    12541 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
    12601 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
    12661 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
    12721 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
    12781 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctata
//
