LOCUS       sequence-257667-        9267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of GSG1-l: C-terminal (amino acid position: STOP 
            codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9267)
  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 9267)
  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 GSG1l-GFP KI
FEATURES             Location/Qualifiers
     source          1..9267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          157..384
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             405..431
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             435..455
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             465..4565
                     /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             465..4565
                     /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             4569..4589
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4629..4646)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4635..4842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4851..4991
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4851..4980
                     /label=AAV2 ITR
     rep_origin      5066..5521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5153..5172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5363..5384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5538..5557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5657..5679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5717..5735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5803..5907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5908..5976,5977..6768)
                     /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(6126..6145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6939..7527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7428..7447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7589..7837
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7589..7609
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7760..7779
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7859..7934
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8079..8081,8082..8795)
                     /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(8079..8081,8082..8792)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8382..8401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8729..8750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        8863..9148
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9150..9267,1..156)
                     /label=chicken beta-actin promoter
ORIGIN
        1 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
       61 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      121 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      181 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      241 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag
      301 aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac
      361 ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catgtaccca tacgatgttc
      421 cagattacgc ttcgccgaag aaaaagcgca aggtcgaagc gtccgacaag aagtacagca
      481 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
      541 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
      601 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
      661 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
      721 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
      781 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
      841 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
      901 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
      961 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     1021 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     1081 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     1141 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     1201 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     1261 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     1321 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     1381 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     1441 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     1501 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     1561 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     1621 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     1681 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     1741 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     1801 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     1861 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     1921 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     1981 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     2041 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     2101 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     2161 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     2221 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     2281 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     2341 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     2401 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     2461 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     2521 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     2581 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     2641 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     2701 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     2761 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     2821 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     2881 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     2941 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     3001 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     3061 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     3121 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     3181 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     3241 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     3301 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     3361 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     3421 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     3481 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     3541 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     3601 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     3661 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     3721 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     3781 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     3841 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     3901 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     3961 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     4021 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     4081 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     4141 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     4201 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     4261 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     4321 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     4381 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     4441 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     4501 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     4561 gcgacagccc caagaagaag agaaaggtgg aggccagcta agaattctaa ctagagctcg
     4621 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4681 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4741 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4801 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
     4861 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4921 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4981 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5041 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5101 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     5161 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5221 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5281 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5341 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     5401 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     5461 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5521 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5581 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5641 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5701 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5761 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5821 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5881 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5941 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6001 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6061 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6121 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6181 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6241 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6301 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6361 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6421 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6481 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6541 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6601 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     6661 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6721 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6781 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6841 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6901 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6961 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7021 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7081 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7141 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7201 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7261 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7321 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7381 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7441 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7501 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7561 gccttttgct ggccttttgc tcacatgtga gggcctattt cccatgattc cttcatattt
     7621 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     7681 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     7741 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     7801 cttggcttta tatatcttgt ggaaaggacg aaacaccgct ggggcactgg gtgtgatggt
     7861 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     7921 caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc
     7981 cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaagcttc cccatcacac
     8041 ccagtgcccc agaaggtagc ggctccagta gatctggggt gagcaagggc gaggagctgt
     8101 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     8161 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     8221 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     8281 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     8341 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     8401 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     8461 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     8521 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     8581 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     8641 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     8701 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     8761 ggatcactct cggcatggac gagctgtaca agtaacccca tcacacccag tgccccagct
     8821 cgaggatcct aggacgcgtc ctaggacgcg tccagaggta cccgttacat aacttacggt
     8881 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc
     8941 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     9001 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     9061 gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     9121 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     9181 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     9241 tgcagcgatg ggggcggggg ggggggg
//
