LOCUS       sequence-259499-        9663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for sgRNAs cloned into the BsaI sites and for 
            expression of Cas9 with 126aa MRN-recruiting domain from HSV-1 UL12 
            fused to C-terminus of Cas9, linked to mCherry via a T2A peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9663)
  AUTHORS   Reuven N, Adler J, Broennimann K, Myers N, Shaul Y
  TITLE     Recruitment of DNA Repair MRN Complex by Intrinsically Disordered 
            Protein Domain Fused to Cas9 Improves Efficiency of CRISPR-Mediated 
            Genome Editing.
  JOURNAL   Biomolecules. 2019 Oct 8;9(10). pii: biom9100584. doi: 
            10.3390/biom9100584.
  PUBMED    31597252
REFERENCE   2  (bases 1 to 9663)
  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: 
            "Biomolecules."; date: "2019-10-8"; volume: "9(10). pii: 
            biom9100584. doi"; pages: " 10.3390/biom9100584"
COMMENT     Sequence Label: pU6-(BsaI)_CBh-UC-Cas9-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..9663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..255
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7..27
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     178..197
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        276..351
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        448..733
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        735..1012
                     /label=chicken beta-actin promoter
     intron          1013..1241
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1253..1262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1262..1327
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1334..1354
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1379..5479
                     /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             1379..5479
                     /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             5909..5962
                     /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             5969..6679
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6114..6132)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(6704..6721)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6710..6917
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6926..7066
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6926..7055
                     /label=AAV2 ITR
     rep_origin      7141..7596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7228..7247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7438..7459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7613..7632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7732..7754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7792..7810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7878..7982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7983..8051,8052..8843)
                     /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(8201..8220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9014..9602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9503..9522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccccacgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
       61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      241 aaaggacgaa acaccggaga cctgtacagg tctctgtttt agagctagaa atagcaagtt
      301 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
      361 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
      421 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
      481 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
      541 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      601 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
      661 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      721 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      781 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      841 gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc
      901 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      961 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     1021 tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     1081 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     1141 gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt
     1201 ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat
     1261 ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga
     1321 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     1381 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
     1441 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     1501 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     1561 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     1621 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     1681 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     1741 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     1801 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     1861 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     1921 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     1981 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     2041 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     2101 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     2161 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     2221 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     2281 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     2341 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     2401 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     2461 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     2521 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     2581 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     2641 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     2701 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     2761 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     2821 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     2881 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     2941 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     3001 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     3061 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     3121 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     3181 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     3241 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     3301 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     3361 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     3421 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     3481 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     3541 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     3601 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     3661 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     3721 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     3781 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     3841 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     3901 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     3961 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     4021 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     4081 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     4141 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     4201 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     4261 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     4321 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     4381 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     4441 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     4501 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     4561 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     4621 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     4681 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     4741 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     4801 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     4861 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     4921 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     4981 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     5041 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     5101 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     5161 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     5221 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     5281 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     5341 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     5401 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     5461 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     5521 gaaaatggag tccacgggag gcccagcatg tccgccggga cgcaccgtga ctaagcgttc
     5581 ctgggccctg gccgaggaca cccctcgtgg ccccgacagc ccccccaagc gcccccgccc
     5641 taacagtctt ccgctaacaa ccaccttccg tcccctgccc cccccacccc agacgacgtc
     5701 agctgtggac ccgagctccc attcgcccgt taacccccca cgtgatcagc acgccaccga
     5761 caccgcagac gaaaagcccc gggccgcgtc gccggcactt tctgacgcct cagggcctcc
     5821 gaccccagac attccgctat ctcctggggg cacccacgcc cgcgacccgg acgccgatcc
     5881 cgactccccg gaccttgact ctaagcttga gggcagagga agtctgctaa catgcggtga
     5941 cgtggaggag aatcccggcc ctgctagcat ggtgagcaag ggcgaggagg ataacatggc
     6001 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     6061 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     6121 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     6181 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     6241 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     6301 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     6361 caccaacttc ccctcagacg gccccgtaat gcagaagaaa accatgggct gggaggcctc
     6421 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     6481 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     6541 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     6601 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     6661 ggacgagctg tacaagtaag aattcctaga gctcgctgat cagcctcgac tgtgccttct
     6721 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     6781 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     6841 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     6901 agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     6961 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     7021 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     7081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     7141 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7201 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7261 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     7321 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     7381 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7441 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     7501 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7561 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     7621 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     7681 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     7741 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     7801 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     7861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     8041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     8101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     8161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     8221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     8281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     8341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     8401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     8461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     8521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     8581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     8641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     8701 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     8761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     8821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     8881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     9001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     9061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     9121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     9181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     9241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     9301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     9361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     9421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     9481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     9541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     9601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     9661 tgt
//
