LOCUS       sequence-343190-        9689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-2A-GFP expression vector bearing two sgRNAs targeting the 
            telomeric human NANOG CTCF-boundary..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9689)
  AUTHORS   Wu HJ, Landshammer A, Stamenova EK, Bolondi A, Kretzmer H, Meissner 
            A, Michor F
  TITLE     Topological isolation of developmental regulators in mammalian 
            genomes.
  JOURNAL   Nat Commun. 2021 Aug 12;12(1):4897. doi: 10.1038/s41467-021-24951-7.
  PUBMED    34385432
REFERENCE   2  (bases 1 to 9689)
  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: "Nat 
            Commun."; date: "2021-08-12"; volume: "12(1):4897. doi"; pages: " 
            10.1038/s41467-021-24951-7"
COMMENT     Sequence Label: 2X_pX458_pSpCas9(BB)-2A-GFP_NANOG_bKO
FEATURES             Location/Qualifiers
     source          1..9689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..265
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     17..37
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_RNA        286..361
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        727..802
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        871..1156
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1422..1649
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1661..1670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1670..1735
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1742..1762
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1787..5887
                     /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             1787..5887
                     /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             5888..5935
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5951..6004
                     /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             join(6005..6007,6008..6718)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6005..6007,6008..6718)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6308..6327)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6655..6676
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6746..6763)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6752..6959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6968..7108
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6968..7097
                     /label=AAV2 ITR
     rep_origin      7183..7638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7270..7289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7480..7501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7655..7674)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7774..7796
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7834..7852)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7920..8024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8025..8093,8094..8885)
                     /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(8243..8262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9056..9644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9545..9564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
       61 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
      121 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      181 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      241 tatcttgtgg aaaggacgaa acacccatcc tttgatacac cccctgtttt agagctagaa
      301 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      361 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      421 tgcgccaatt ctgcagacaa atggctctag aggtaccaat atgcattttc ccatgattcc
      481 ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt
      541 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
      601 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
      661 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacacctgtt agtctttgtt
      721 caaccggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      781 aaaagtggca ccgagtcggt gcttttttgt tttcctgcag gagatttagc gcgtgcgcca
      841 attctgcaga caaatggctc tagaggtacc cgttacataa cttacggtaa atggcccgcc
      901 tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt
      961 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1021 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1081 ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1141 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     1201 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     1261 ggcggggggg gggggggggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
     1321 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
     1381 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     1441 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     1501 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagctga
     1561 gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt aattacctgg
     1621 agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg actataagga
     1681 ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc
     1741 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
     1801 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     1861 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     1921 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     1981 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     2041 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     2101 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     2161 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     2221 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     2281 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     2341 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     2401 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     2461 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     2521 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     2581 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     2641 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     2701 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     2761 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     2821 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     2881 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     2941 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     3001 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     3061 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     3121 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     3181 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     3241 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     3301 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     3361 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     3421 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     3481 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     3541 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     3601 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     3661 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     3721 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     3781 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     3841 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     3901 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     3961 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     4021 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     4081 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     4141 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     4201 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     4261 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     4321 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     4381 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     4441 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     4501 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     4561 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     4621 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     4681 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     4741 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     4801 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     4861 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     4921 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     4981 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     5041 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     5101 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     5161 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     5221 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     5281 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     5341 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     5401 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     5461 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     5521 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     5581 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     5641 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     5701 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     5761 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     5821 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     5881 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt
     5941 cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     6001 cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     6061 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     6121 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     6181 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     6241 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     6301 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     6361 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     6421 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     6481 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     6541 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     6601 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     6661 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagga
     6721 attctaacta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6781 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6841 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6901 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
     6961 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     7021 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     7081 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     7141 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     7201 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     7261 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7321 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7381 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7441 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7501 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     7561 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     7621 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7681 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7741 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7801 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     7861 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     7921 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7981 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8041 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8101 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8161 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8221 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8281 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8341 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8401 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8461 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8521 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8581 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8641 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8701 gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag
     8761 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8821 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8881 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8941 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9001 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9661 cctttttacg gttcctggcc ttttgctgg
//
