LOCUS       sequence-304490-        9345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SpyCas9, puromycin resistance and 
            human Polb guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9345)
  AUTHORS   Danner E, Lebedin M, de la Rosa K, Kuhn R
  TITLE     A homology independent sequence replacement strategy in human cells 
            using a CRISPR nuclease.
  JOURNAL   Open Biol. 2021 Jan;11(1):200283. doi: 10.1098/rsob.200283. Epub 
            2021 Jan 27.
  PUBMED    33499763
REFERENCE   2  (bases 1 to 9345)
  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: "Open 
            Biol."; date: "2021-01"; volume: "11(1):200283. doi"; pages: " 
            10.1098/rsob.200283. Epub 2021 Jan 27"
COMMENT     Sequence Label: pCbh-SpyCas9-T2A-puro-pA_U6-sgPolb (pE047)
FEATURES             Location/Qualifiers
     source          1..9345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..4123
                     /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             23..4123
                     /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             4124..4171
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4187..4240
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4241..4840
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4241..4260)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4737..4757
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4868..4885)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4874..5081
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5090..5219
                     /label=AAV2 ITR
     primer_bind     complement(5293..5311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5379..5483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5484..5552,5553..6344)
                     /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(5702..5721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6515..7103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7004..7023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7165..7413
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7432..7507
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7600..7848
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7887..7905
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        7957..7975
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        8022..8270
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8309..8327
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        8379..8397
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     enhancer        8438..8723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        8725..9002
                     /label=chicken beta-actin promoter
     intron          9003..9230
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      9242..9251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9251..9316
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             9323..9343
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
       61 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
      121 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
      181 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
      241 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
      301 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
      361 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
      421 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
      481 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
      541 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
      601 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
      661 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
      721 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
      781 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
      841 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
      901 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
      961 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     1021 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     1081 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     1141 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     1201 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     1261 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     1321 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     1381 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     1441 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     1501 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     1561 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     1621 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     1681 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     1741 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     1801 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     1861 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     1921 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     1981 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     2041 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     2101 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     2161 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     2221 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     2281 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     2341 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     2401 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     2461 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     2521 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     2581 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     2641 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     2701 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     2761 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     2821 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     2881 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     2941 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     3001 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     3061 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     3121 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     3181 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     3241 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     3301 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     3361 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     3421 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     3481 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     3541 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     3601 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     3661 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     3721 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     3781 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     3841 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     3901 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     3961 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     4021 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     4081 cctgtacgag acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac
     4141 gaaaaaggcc ggccaggcaa aaaagaaaaa ggaattcggc agtggagagg gcagaggaag
     4201 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca atgaccgagt acaagcccac
     4261 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4321 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4381 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4441 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4501 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4561 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4621 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4681 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4741 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4801 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gaattctaac tagagctcgc
     4861 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4921 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4981 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5041 aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta
     5101 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     5161 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     5221 tgcctgcaat cactacattc acatctgatt tcagtagttc cttacgagag ttcctccgcg
     5281 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5341 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5401 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5461 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5521 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5581 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5641 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5701 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5761 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5821 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5881 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5941 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6001 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6061 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6181 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     6241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7141 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
     7201 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     7261 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     7321 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     7381 gctttatata tcttgtggaa aggacgaaac accgggtctt cgagaagacc tgttttagag
     7441 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     7501 tcggtgcttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt
     7561 agcgcgtgcg ccaattctgc agacaaatgg ctctagagcg agggcctatt tcccatgatt
     7621 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
     7681 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     7741 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     7801 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt aattttgtgt
     7861 gggtcacccg ttttagagct aggccaacat gaggatcacc catgtctgca gggcctagca
     7921 agttaaaata aggctagtcc gttatcaact tggccaacat gaggatcacc catgtctgca
     7981 gggccaagtg gcaccgagtc ggtgcttttt ttgagatatc cgagggccta tttcccatga
     8041 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga
     8101 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     8161 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     8221 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gtgaaaccag
     8281 tttggttacc cgttttagag ctaggccaac atgaggatca cccatgtctg cagggcctag
     8341 caagttaaaa taaggctagt ccgttatcaa cttggccaac atgaggatca cccatgtctg
     8401 cagggccaag tggcaccgag tcggtgcttt ttttgcccgt tacataactt acggtaaatg
     8461 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
     8521 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     8581 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     8641 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     8701 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     8761 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     8821 cgatgggggc gggggggggg ggggggcgcg cgccrggggg gggggggggg ggsggggggg
     8881 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     8941 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     9001 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     9061 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     9121 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
     9181 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
     9241 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
     9301 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcgg
//
