LOCUS       sequence-257671-        9248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of Shank2: C-terminal (amino acid position: STOP 
            codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9248)
  AUTHORS   Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE, 
            Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD
  TITLE     ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope 
            tagging of endogenous proteins in neurons.
  JOURNAL   PLoS Biol. 2020 Apr 10;18(4):e3000665. doi: 
            10.1371/journal.pbio.3000665.
  PUBMED    32275651
REFERENCE   2  (bases 1 to 9248)
  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"; journalName: "PLoS 
            Biol."; date: "2020-04-10"; volume: "18(4):e3000665. doi"; pages: " 
            10.1371/journal.pbio.3000665"
COMMENT     Sequence Label: pORANGE Shank2-GFP KI
FEATURES             Location/Qualifiers
     source          1..9248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          228..455
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             476..502
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             506..526
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             536..4636
                     /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             536..4636
                     /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             4640..4660
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4700..4717)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4706..4913
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4922..5062
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4922..5051
                     /label=AAV2 ITR
     rep_origin      5137..5592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5224..5243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5434..5455
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5609..5628)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5728..5750
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5788..5806)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5874..5978
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5979..6047,6048..6839)
                     /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(6197..6216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7010..7598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7499..7518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7660..7908
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7660..7680
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7831..7850
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7930..8005
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8125..8127,8128..8841)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8125..8127,8128..8838)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8428..8447)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8775..8796
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        8909..9194
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9196..9248,1..227)
                     /label=chicken beta-actin promoter
ORIGIN
        1 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
       61 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
      121 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
      181 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
      241 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      301 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      361 agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt
      421 acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatgtaccc
      481 atacgatgtt ccagattacg cttcgccgaa gaaaaagcgc aaggtcgaag cgtccgacaa
      541 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
      601 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
      661 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
      721 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
      781 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
      841 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
      901 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
      961 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     1021 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     1081 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     1141 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     1201 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     1261 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     1321 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     1381 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     1441 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     1501 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     1561 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     1621 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     1681 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     1741 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     1801 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     1861 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     1921 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     1981 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     2041 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     2101 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     2161 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     2221 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     2281 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     2341 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     2401 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     2461 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     2521 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     2581 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     2641 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     2701 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     2761 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     2821 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     2881 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     2941 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     3001 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     3061 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     3121 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     3181 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     3241 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     3301 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     3361 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     3421 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     3481 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     3541 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     3601 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     3661 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     3721 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     3781 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     3841 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     3901 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     3961 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     4021 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     4081 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     4141 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     4201 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     4261 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     4321 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     4381 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     4441 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     4501 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     4561 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     4621 tcagctggga ggcgacagcc ccaagaagaa gagaaaggtg gaggccagct aagaattcta
     4681 actagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4741 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4801 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4861 gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg
     4921 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4981 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5041 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5101 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     5161 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     5221 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5281 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5341 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5401 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5461 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     5521 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5581 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5641 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5701 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5761 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5821 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5881 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5941 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6001 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6061 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6121 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6181 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6241 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6301 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6361 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6421 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6481 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6541 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6601 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6661 ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg
     6721 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6781 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6841 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6901 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6961 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7021 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7081 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7141 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7201 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7261 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7321 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7381 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7441 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7501 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7561 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7621 tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt tcccatgatt
     7681 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
     7741 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     7801 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     7861 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccga acagctgctg
     7921 gacagataag ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     7981 tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa
     8041 taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaagctt
     8101 cccttatctg tccagcagct gttagtgagc aagggcgagg agctgttcac cggggtggtg
     8161 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     8221 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     8281 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     8341 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     8401 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     8461 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     8521 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     8581 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     8641 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     8701 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     8761 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     8821 atggacgagc tgtacaagta acccttatct gtccagcagc tgttctcgag gatcctagga
     8881 cgcgtcctag gacgcgtcca gaggtacccg ttacataact tacggtaaat ggcccgcctg
     8941 gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata gggactttcc
     9001 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     9061 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     9121 gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     9181 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     9241 cctcccca
//
