LOCUS       sequence-345081-        8403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of spCas9-nickase and one sgRNA targeting 
            the mouse WAPL locus. Use together with pX335-NQL002-WAPL-sgRNA1 
            targeting construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8403)
  AUTHORS   Liu NQ, Maresca M, van den Brand T, Braccioli L, Schijns MMGA, 
            Teunissen H, Bruneau BG, Nora EP, de Wit E
  TITLE     WAPL maintains a cohesin loading cycle to preserve 
            cell-type-specific distal gene regulation.
  JOURNAL   Nat Genet. 2021 Jan;53(1):100-109. doi: 10.1038/s41588-020-00744-4. 
            Epub 2020 Dec 14.
  PUBMED    33318687
REFERENCE   2  (bases 1 to 8403)
  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"; doi: 
            "10.1038/s41588-020-00744-4"; journalName: "Nat Genet"; date: 
            "2021-01"; volume: "53"; issue: "1"; pages: "100-109"
COMMENT     Sequence Label: pX335-NQL003-WAPL-sgRNA2
FEATURES             Location/Qualifiers
     source          1..8403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        443..728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          975..1202
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1223..1249
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1253..1273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1283..1306,1307..1309,1310..5383)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1283..5383
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5387..5407
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5444..5461)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5450..5657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5666..5806
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5666..5795
                     /label=AAV2 ITR
     rep_origin      5881..6336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5968..5987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6178..6199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6353..6372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6472..6494
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6532..6550)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6618..6722
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6723..6791,6792..7583)
                     /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(6941..6960)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7754..8342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8243..8262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ttacctttgc ttcaggtgct gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
      481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
      541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
      661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
      781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
      841 gggggggggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
      901 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      961 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
     1021 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
     1081 acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt
     1141 tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc
     1201 aggttggacc ggtgccacca tgtacccata cgatgttcca gattacgctt cgccgaagaa
     1261 aaagcgcaag gtcgaagcgt ccgacaagaa gtacagcatc ggcctggcca tcggcaccaa
     1321 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     1381 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     1441 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     1501 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     1561 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     1621 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     1681 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     1741 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     1801 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     1861 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     1921 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     1981 gaatggcctg ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     2041 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     2101 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     2161 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     2221 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     2281 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     2341 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     2401 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     2461 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     2521 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     2581 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     2641 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     2701 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     2761 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     2821 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     2881 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     2941 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     3001 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     3061 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     3121 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     3181 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     3241 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     3301 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     3361 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     3421 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     3481 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     3541 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     3601 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     3661 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     3721 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     3781 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     3841 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     3901 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     3961 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     4021 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     4081 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     4141 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     4201 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     4261 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     4321 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     4381 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     4441 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     4501 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     4561 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     4621 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     4681 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     4741 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     4801 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     4861 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     4921 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     4981 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     5041 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     5101 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     5161 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     5221 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     5281 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     5341 cctgtacgag acacggatcg acctgtctca gctgggaggc gacagcccca agaagaagag
     5401 aaaggtggag gccagctaag aattcctaga gctcgctgat cagcctcgac tgtgccttct
     5461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     5641 agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     5701 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     5761 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     5821 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5881 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5941 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6001 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6061 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     6121 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6181 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     6241 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     6301 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     6361 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     6421 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     6481 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     6541 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6601 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6661 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6721 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6781 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6841 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6901 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6961 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7021 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7081 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7141 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7201 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7261 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7321 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7381 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     7441 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7501 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7561 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7621 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7681 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7741 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7801 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7861 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7921 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7981 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8041 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8101 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8161 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8221 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8281 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8341 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8401 tgt
//
