LOCUS       sequence-257607-        9258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of Clathrin light chain alpha: N-terminal (amino 
            acid position: M1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9258)
  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 9258)
  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 GFP-Clta KI
FEATURES             Location/Qualifiers
     source          1..9258
                     /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        330..405
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(534..536,537..1247)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(534..536,537..1247)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(837..856)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1184..1205
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1323..1608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1610..1887
                     /label=chicken beta-actin promoter
     intron          1888..2115
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2136..2162
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2166..2186
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2196..6296
                     /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             2196..6296
                     /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             6300..6320
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6360..6377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6366..6573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6582..6722
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6582..6711
                     /label=AAV2 ITR
     rep_origin      6797..7252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6884..6903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7094..7115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7269..7288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7388..7410
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7448..7466)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7534..7638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7639..7707,7708..8499)
                     /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(7857..7876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8670..9258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9159..9178
                     /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 ggatccaact cagccatgag ttttagagct agaaatagca agttaaaata
      361 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
      421 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
      481 acaaatggct ctagaagctt ggatccaact cagccatgag ggcaggtagc ggagtgagca
      541 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      601 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      661 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      721 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      781 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      841 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      901 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      961 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1021 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1081 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1141 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1201 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt agcggaggat
     1261 ccaactcagc catgagggct cgaggatcct aggacgcgtc ctaggacgcg tccagaggta
     1321 cccgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1381 ttgacgtcaa tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1441 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1501 gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc agtacatgac cttatgggac
     1561 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
     1621 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
     1681 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca
     1741 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
     1801 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     1861 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     1921 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1981 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg
     2041 tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa
     2101 atcacttttt ttcaggttgg accggtgcca ccatgtaccc atacgatgtt ccagattacg
     2161 cttcgccgaa gaaaaagcgc aaggtcgaag cgtccgacaa gaagtacagc atcggcctgg
     2221 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     2281 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     2341 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     2401 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     2461 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     2521 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     2581 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     2641 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     2701 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     2761 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     2821 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     2881 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     2941 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     3001 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     3061 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     3121 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     3181 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     3241 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     3301 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     3361 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     3421 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     3481 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     3541 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     3601 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     3661 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     3721 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     3781 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     3841 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     3901 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     3961 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     4021 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     4081 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     4141 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     4201 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     4261 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     4321 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     4381 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     4441 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     4501 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     4561 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     4621 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     4681 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     4741 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     4801 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     4861 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     4921 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     4981 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     5041 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     5101 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     5161 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     5221 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     5281 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     5341 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     5401 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     5461 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     5521 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     5581 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     5641 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     5701 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     5761 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     5821 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     5881 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     5941 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     6001 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     6061 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     6121 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     6181 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     6241 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacagcc
     6301 ccaagaagaa gagaaaggtg gaggccagct aagaattcta actagagctc gctgatcagc
     6361 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6421 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     6481 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     6541 ggattgggaa gagaatagca ggcatgctgg ggagcggccg caggaacccc tagtgatgga
     6601 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     6661 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     6721 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     6781 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6841 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     6901 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6961 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     7021 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7081 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     7141 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     7201 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     7261 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     7321 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     7381 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     7441 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     7501 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7861 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     8041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     8101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     8161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     8221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     8281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8341 gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     9001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     9061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     9121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     9181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     9241 gggcggagcc tatggaaa
//
