LOCUS       sequence-494281-        9210 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  CRISPR vector co-expressing Cas9 and a mouse Gpr124 gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9210)
  AUTHORS   Heiden R, Hannig L, Kuo CJ, Ergun S, Braunger BM, Vallon M
  TITLE     WNT7A/B assemble a GPR124-RECK-LRP5/6 co-receptor complex to 
            activate beta-catenin signaling in brain endothelial cells.
  JOURNAL   J Biol Chem. 2025 Sep 4:110682. doi: 10.1016/j.jbc.2025.110682.
  PUBMED    40914247
REFERENCE   2  (bases 1 to 9210)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2025 Sep 4:110682. doi: 10.1016/j.jbc.2025.110682."
COMMENT     Sequence Label: pX459-Gpr124 gRNA
FEATURES             Location/Qualifiers
     source          1..9210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        443..728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1041..1268
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1280..1289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1289..1354
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1361..1381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1406..5506
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1406..5506
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5507..5554
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5570..5623
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5624..6223
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5624..5643)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6120..6140
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6251..6268)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6257..6464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6473..6613
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6473..6602
                     /label=AAV2 ITR
     rep_origin      6688..7143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6775..6794)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6985..7006
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7160..7179)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7279..7301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7339..7357)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7425..7529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7530..7598,7599..8390)
                     /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     complement(7748..7767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8561..9149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9050..9069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg agcacttgca accgcggata gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
      481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
      541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
      661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
      781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
      841 gggggggggg ggcgccccct gggggggggg gggcgggggg gggggggggg gggggggggg
      901 gggggggggg cggggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      961 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     1021 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
     1081 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     1141 ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg
     1201 gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt
     1261 tttttcaggt tggaccggtg ccaccatgga ctataaggac cacgacggag actacaagga
     1321 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
     1381 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
     1441 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     1501 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     1561 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     1621 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     1681 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     1741 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     1801 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     1861 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     1921 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     1981 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     2041 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     2101 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     2161 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     2221 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     2281 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     2341 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     2401 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     2461 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     2521 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     2581 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     2641 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     2701 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     2761 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     2821 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     2881 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     2941 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     3001 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     3061 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     3121 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     3181 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     3241 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     3301 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     3361 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     3421 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     3481 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     3541 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     3601 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     3661 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     3721 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     3781 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     3841 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     3901 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     3961 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     4021 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     4081 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     4141 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     4201 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     4261 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     4321 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     4381 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     4441 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     4501 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     4561 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     4621 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     4681 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     4741 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     4801 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     4861 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     4921 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     4981 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     5041 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     5101 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     5161 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     5221 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     5281 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     5341 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     5401 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     5461 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
     5521 cacgaaaaag gccggccagg caaaaaagaa aaaggaattc ggcagtggag agggcagagg
     5581 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaatgaccg agtacaagcc
     5641 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     5701 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     5761 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5821 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5881 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5941 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     6001 caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     6061 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     6121 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6181 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagaattct aactagagct
     6241 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     6301 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     6361 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     6421 agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc
     6481 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     6541 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     6601 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6661 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     6721 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     6781 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6841 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6901 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6961 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     7021 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     7081 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     7141 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     7201 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7261 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7321 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     7381 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     7441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7501 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7561 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7621 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7681 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7741 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7801 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7861 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7921 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7981 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8041 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     8101 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     8161 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     8221 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     8281 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8341 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     8401 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     8461 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8521 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     8581 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     8641 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     8701 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8761 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8821 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8881 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8941 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9001 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9061 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9121 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     9181 tggccttttg ctggcctttt gctcacatgt
//
