LOCUS       sequence-327132-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insertion of NIPBL degron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8510)
  AUTHORS   Zhu Y, Denholtz M, Lu H, Murre C
  TITLE     Calcium signaling instructs NIPBL recruitment at active enhancers 
            and promoters via distinct mechanisms to reconstruct genome 
            compartmentalization.
  JOURNAL   Genes Dev. 2021 Jan 1;35(1-2):65-81. doi: 10.1101/gad.343475.120. 
            Epub 2020 Dec 17.
  PUBMED    33334824
REFERENCE   2  (bases 1 to 8510)
  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: "Genes 
            Dev."; date: "2021-01-1"; volume: "35(1-2):65-81. doi"; pages: " 
            10.1101/gad.343475.120. Epub 2020 Dec 17"
COMMENT     Sequence Label: px330 NIPBL gRNA
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          97..324
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      336..345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             345..410
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             417..437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             462..4562
                     /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             462..4562
                     /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             4563..4610
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4638..4655)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4644..4851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4860..5000
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4860..4989
                     /label=AAV2 ITR
     rep_origin      5075..5530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5162..5181)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5372..5393
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5547..5566)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5666..5688
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5726..5744)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5812..5916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5917..5985,5986..6777)
                     /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(6135..6154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6948..7536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7437..7456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7598..7846
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7598..7618
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7769..7788
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7868..7943
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8040..8325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8327..8510,1..96)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
       61 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      121 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      181 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag
      241 aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac
      301 ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat aaggaccacg
      361 acggagacta caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa
      421 agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag tacagcatcg
      481 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
      541 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
      601 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
      661 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
      721 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
      781 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
      841 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
      901 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
      961 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     1021 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     1081 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     1141 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
     1201 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     1261 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     1321 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     1381 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     1441 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     1501 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     1561 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     1621 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     1681 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     1741 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     1801 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     1861 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     1921 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     1981 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     2041 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     2101 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     2161 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     2221 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     2281 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     2341 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     2401 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     2461 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     2521 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     2581 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     2641 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     2701 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     2761 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     2821 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     2881 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     2941 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga
     3001 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     3061 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     3121 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     3181 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     3241 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     3301 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     3361 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     3421 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     3481 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     3541 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     3601 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     3661 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
     3721 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     3781 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
     3841 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
     3901 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     3961 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     4021 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     4081 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
     4141 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     4201 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
     4261 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     4321 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
     4381 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     4441 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     4501 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
     4561 acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag taagaattcc
     4621 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4681 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4741 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4801 gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca
     4861 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4921 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4981 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5041 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     5101 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     5161 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5221 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5281 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5341 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5401 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     5461 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5521 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5581 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5641 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5701 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5761 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5881 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5941 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6001 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6061 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6121 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6601 attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg
     6661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7081 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7561 cggttcctgg ccttttgctg gccttttgct cacatgtgag ggcctatttc ccatgattcc
     7621 ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt
     7681 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     7741 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     7801 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcca gaacttcagg
     7861 atgaatggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     7921 aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata
     7981 aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agaggtaccc
     8041 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     8101 acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     8161 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     8221 gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt atgggacttt
     8281 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc
     8341 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     8401 attttttaat tattttgtgc agcgatgggg gcgggggggg gggggggggg cgcgcgccag
     8461 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
//
