LOCUS       sequence-257602-        9258 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Endogenous tagging of Complexin1: C-terminal (amino acid position: 
            L128).
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 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 Cplx1-GFP KI
FEATURES             Location/Qualifiers
     source          1..9258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          135..362
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             383..409
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             413..433
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             443..4543
                     /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             443..4543
                     /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             4547..4567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4607..4624)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4613..4820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4829..4969
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4829..4958
                     /label=AAV2 ITR
     rep_origin      5044..5499
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5131..5150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5341..5362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5516..5535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5635..5657
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5695..5713)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5781..5885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5886..5954,5955..6746)
                     /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(6104..6123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6917..7505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7406..7425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7567..7815
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7567..7587
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7738..7757
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7837..7912
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8040..8042,8043..8756)
                     /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(8040..8042,8043..8753)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8343..8362)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8690..8711
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        8832..9117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
       61 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      121 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      181 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      241 acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt
      301 tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc
      361 aggttggacc ggtgccacca tgtacccata cgatgttcca gattacgctt cgccgaagaa
      421 aaagcgcaag gtcgaagcgt ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
      481 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
      541 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
      601 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
      661 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
      721 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
      781 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
      841 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
      901 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
      961 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     1021 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     1081 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     1141 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     1201 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     1261 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     1321 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     1381 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     1441 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     1501 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     1561 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     1621 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     1681 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     1741 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     1801 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     1861 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     1921 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     1981 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     2041 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     2101 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     2161 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     2221 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     2281 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     2341 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     2401 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     2461 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     2521 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     2581 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     2641 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     2701 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     2761 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     2821 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     2881 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     2941 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     3001 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     3061 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     3121 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     3181 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     3241 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     3301 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     3361 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     3421 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     3481 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     3541 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     3601 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     3661 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     3721 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     3781 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     3841 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     3901 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     3961 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     4021 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     4081 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     4141 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     4201 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     4261 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     4321 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     4381 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     4441 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     4501 cctgtacgag acacggatcg acctgtctca gctgggaggc gacagcccca agaagaagag
     4561 aaaggtggag gccagctaag aattctaact agagctcgct gatcagcctc gactgtgcct
     4621 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4681 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4741 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     4801 aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4861 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4921 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4981 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5041 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5101 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5161 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5221 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5281 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5341 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     5401 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     5461 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5521 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5581 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5641 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5701 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5761 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5821 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5881 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5941 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6001 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6061 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6601 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7561 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     7621 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     7681 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     7741 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     7801 aaaggacgaa acaccggtac ctgcctgggc cactgcgttt tagagctaga aatagcaagt
     7861 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt
     7921 tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat
     7981 tctgcagaca aatggctcta gaagcttccc gcagtggccc aggcaggtac agcggtggag
     8041 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     8101 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     8161 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     8221 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     8281 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     8341 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     8401 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     8461 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     8521 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     8581 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     8641 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     8701 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtgagcgg
     8761 tggacccgca gtggcccagg caggtacctc gaggatccta ggacgcgtcc taggacgcgt
     8821 ccagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     8881 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
     8941 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     9001 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
     9061 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     9121 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     9181 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggnnnn
     9241 cgccaggcgg ggcggggc
//
