LOCUS       sequence-383803-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sequence specific sgRNA that guide Cas9 to the genomic region 
            encoding the bovine Creb5 DNA binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Zhang CH, Gao Y, Jadhav U, Hung HH, Holton KM, Grodzinsky AJ, 
            Shivdasani RA, Lassar AB
  TITLE     Creb5 establishes the competence for Prg4 expression in articular 
            cartilage.
  JOURNAL   Commun Biol. 2021 Mar 12;4(1):332. doi: 10.1038/s42003-021-01857-0.
  PUBMED    33712729
REFERENCE   2  (bases 1 to 13013)
  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: "Commun 
            Biol."; date: "2021-03-12"; volume: "4(1):332. doi"; pages: " 
            10.1038/s42003-021-01857-0"
COMMENT     Sequence Label: LentiCRISPRCreb5gRNA
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        85..414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      265..400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     327..346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        462..509
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             528..902
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1032..1153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1069..1088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1123..1142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1202..1218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1202..1218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1215..1237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1226..1242
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1250..1256,1257..1274,1275..1280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1295..1316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1433..1450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1604..2192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1684..1703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2363..3154,3155..3223))
                     /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     2986..3005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3224..3328)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3403..3422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3594..3973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3975..4173
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4128..4148
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4191..4371
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4418..4543
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5036..5269
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5454..5498
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5796..5913
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5964..6212
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5964..5984
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6135..6154
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6234..6309
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6371..6582
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6601..6610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6607..10710
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6607..10710
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             10711..10758
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10759..10782
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10792..10848
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             10849..11445
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     11342..11362
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11461..12049
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11514..11534)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11932..11943)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12121..12354
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12380..12397)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12386..12610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12656..13013,1..71)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12743..12762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12953..12974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
       61 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
      181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      241 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      301 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
      361 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
      421 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
      481 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
      541 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
      601 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
      661 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
      721 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
      781 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
      841 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
      901 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
      961 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     1021 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1081 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1141 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     1201 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1261 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1321 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1381 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3301 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     3361 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     3421 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     3481 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     3541 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     3601 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     3661 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3721 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3781 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3841 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3901 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3961 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     4021 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     4081 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     4141 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
     4201 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4261 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4321 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     4381 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     4441 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     4501 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     4561 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     4621 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     4681 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     4741 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     4801 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     4861 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     4921 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     4981 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     5041 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     5101 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     5161 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     5221 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     5281 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     5341 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     5401 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     5461 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     5521 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     5581 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     5641 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     5701 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     5761 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     5821 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     5881 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     5941 ccagtttggt taattaaggt accgagggcc tatttcccat gattccttca tatttgcata
     6001 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     6061 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     6121 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     6181 ctttatatat cttgtggaaa ggacgaaaca ccgctgaagc tgcatgtttg tctgttttag
     6241 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     6301 agtcggtgct tttttgaatt cgctagctag gtcttgaaag gagtgggaat tggctccggt
     6361 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     6421 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     6481 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     6541 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggaccggtt ctagagcgct
     6601 gccaccatgg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     6661 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     6721 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     6781 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     6841 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     6901 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     6961 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     7021 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     7081 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     7141 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     7201 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     7261 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     7321 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     7381 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     7441 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     7501 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     7561 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     7621 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     7681 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     7741 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     7801 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     7861 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     7921 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     7981 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     8041 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     8101 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     8161 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     8221 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     8281 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     8341 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     8401 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     8461 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     8521 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     8581 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     8641 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     8701 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     8761 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     8821 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     8881 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     8941 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     9001 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     9061 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     9121 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     9181 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     9241 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     9301 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     9361 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     9421 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     9481 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     9541 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     9601 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     9661 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     9721 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     9781 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     9841 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     9901 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     9961 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
    10021 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
    10081 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
    10141 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
    10201 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
    10261 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
    10321 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
    10381 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
    10441 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
    10501 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
    10561 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
    10621 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
    10681 cggatcgacc tgtctcagct gggaggcgac aagcgacctg ccgccacaaa gaaggctgga
    10741 caggctaaga agaagaaaga ttacaaagac gatgacgata agggatccgg cgcaacaaac
    10801 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgac cgagtacaag
    10861 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
    10921 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
    10981 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
    11041 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
    11101 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
    11161 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
    11221 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
    11281 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
    11341 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
    11401 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
    11461 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
    11521 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
    11581 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
    11641 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
    11701 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
    11761 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
    11821 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
    11881 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
    11941 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
    12001 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
    12061 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
    12121 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
    12181 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
    12241 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
    12301 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
    12361 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
    12421 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
    12481 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
    12541 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
    12601 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
    12661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
    12721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    12781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    12841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    12901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    12961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga ttt
//
