LOCUS       sequence-257256-        9223 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Endogenous tagging of CaMKIIa: N-terminal (amino acid position: 
            before startcodon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9223)
  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 9223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 mEos3.2-Camk2a KI
FEATURES             Location/Qualifiers
     source          1..9223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     62..82
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     233..252
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        332..407
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             541..1218
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     enhancer        1288..1573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1575..1852
                     /label=chicken beta-actin promoter
     intron          1853..2080
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2101..2127
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2131..2151
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2161..6261
                     /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             2161..6261
                     /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             6265..6285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6325..6342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6331..6538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6547..6687
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6547..6676
                     /label=AAV2 ITR
     rep_origin      6762..7217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6849..6868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7059..7080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7234..7253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7353..7375
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7413..7431)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7499..7603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7604..7672,7673..8464)
                     /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(7822..7841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8635..9223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9124..9143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      121 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      301 acgaaacacc gctgcctgcc cagtgccagg agttttagag ctagaaatag caagttaaaa
      361 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc
      481 agacaaatgg ctctagaagc ttgctagacc ctcctggcac tgggcaggca ggctagcagg
      541 atgagtgcga ttaagccaga catgaagatc aaactccgta tggaaggcaa cgtaaacggg
      601 caccactttg tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa acagagtatg
      661 gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct gaccactgca
      721 ttccattacg gcaacagggt attcgccaaa tatccagaca acatacaaga ctattttaag
      781 cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga cgggggcatt
      841 tgcaacgcca gaaacgacat aacaatggaa ggggacactt tctataataa agttcgattt
      901 tatggtacca actttcccgc caatggtcca gttatgcaga agaagacgct gaaatgggag
      961 ccctccactg agaaaatgta tgtgcgtgat ggagtgctga cgggtgatat tgagatggct
     1021 ttgttgcttg aaggaaatgc ccattaccga tgtgacttca gaactactta caaagctaag
     1081 gagaagggtg tcaagttacc aggcgcccac tttgtggacc actgcattga gattttaagc
     1141 catgacaaag attacaacaa ggttaagctg tatgagcatg ctgttgctca ttctggattg
     1201 cctgacaatg ccagacgagc tcgagccctc ctggcactgg gcaggcaggg atcctaggac
     1261 gcgtcctagg acgcgtccag aggtacccgt tacataactt acggtaaatg gcccgcctgg
     1321 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
     1381 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     1441 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
     1501 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1561 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     1621 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     1681 gggggggggg ggggggcgcg cgccgggggg gggggggggg ggsggggggg ggggsggggs
     1741 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     1801 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     1861 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     1921 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     1981 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
     2041 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
     2101 tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt cgaagcgtcc
     2161 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     2221 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     2281 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     2341 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     2401 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     2461 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     2521 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     2581 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     2641 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     2701 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     2761 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     2821 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     2881 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     2941 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3001 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3061 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3121 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     3181 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     3241 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     3301 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     3361 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     3421 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     3481 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     3541 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     3601 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     3661 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     3721 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     3781 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     3841 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     3901 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     3961 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4021 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4081 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4141 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     4201 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     4261 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     4321 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     4381 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     4441 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     4501 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     4561 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     4621 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     4681 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     4741 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     4801 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     4861 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     4921 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     4981 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5041 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5101 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     5161 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     5221 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     5281 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     5341 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     5401 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     5461 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     5521 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     5581 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     5641 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     5701 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     5761 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     5821 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     5881 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     5941 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6001 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6061 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6121 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     6181 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     6241 ctgtctcagc tgggaggcga cagccccaag aagaagagaa aggtggaggc cagctaagaa
     6301 ttctaactag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6481 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
     6541 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     6601 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     6661 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     6721 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     6781 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     6841 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6901 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6961 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7021 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7081 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     7141 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     7201 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7261 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7321 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7381 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7441 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7501 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7561 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7621 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7681 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7741 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7801 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7861 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7921 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7981 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8041 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8101 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8161 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8221 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8281 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     8341 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8401 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8461 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8761 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     9001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaa
//
