LOCUS       sequence-488029-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one CRISPRko plasmid containing Cas9 and guide RNA targeting 
            FXN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Ast T, Meisel JD, Patra S, Wang H, Grange RMH, Kim SH, Calvo SE, 
            Orefice LL, Nagashima F, Ichinose F, Zapol WM, Ruvkun G, Barondeau 
            DP, Mootha VK
  TITLE     Hypoxia Rescues Frataxin Loss by Restoring Iron Sulfur Cluster 
            Biogenesis.
  JOURNAL   Cell. 2019 May 30;177(6):1507-1521.e16. doi: 
            10.1016/j.cell.2019.03.045. Epub 2019 Apr 25.
  PUBMED    31031004
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"; doi: 
            "10.1016/j.cell.2019.03.045"; journalName: "Cell"; date: 
            "2019-05-30- 30"; volume: "177"; issue: "6"; pages: "1507-1521.e16"
COMMENT     Sequence Label: pLentiCRISPRv2-sgFXN-2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1211..1230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1402..1781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1783..1981
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1936..1956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1999..2179
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2226..2351
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2844..3077
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3262..3306
                     /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    3604..3721
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3772..4020
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3772..3792
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3943..3962
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4042..4117
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4179..4390
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4409..4418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4415..8518
                     /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             4415..8518
                     /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             8519..8566
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8567..8590
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8600..8656
                     /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             8657..9253
                     /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     9150..9170
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9269..9857
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9322..9342)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9740..9751)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9929..10162
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10188..10205)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10194..10418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10464..10892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10551..10570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10761..10782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10901..10921)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10906..11235
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11086..11221
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11148..11167
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11283..11330
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11349..11723
                     /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    11853..11974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11890..11909)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11944..11963
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12023..12039)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12023..12039)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12036..12058)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12047..12063
                     /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(12071..12077,12078..12095,12096..12101))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12116..12137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12254..12271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12425..13013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12505..12524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     1381 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     1441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1741 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1981 gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     2041 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2101 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2161 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     2221 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     2281 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     2341 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     2401 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     2461 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     2521 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     2581 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     2641 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     2701 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     2761 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     2821 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     2881 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     2941 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     3001 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     3061 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     3121 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     3181 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     3241 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     3301 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     3361 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     3421 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     3481 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     3541 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
     3601 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     3661 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     3721 tcgggtttat tacagggaca gcagagatcc agtttggtta attaaggtac cgagggccta
     3781 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga
     3841 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     3901 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     3961 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
     4021 gccaggccca gaccctcacc cgttttagag ctagaaatag caagttaaaa taaggctagt
     4081 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgaattcg ctagctaggt
     4141 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     4201 agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg
     4261 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     4321 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     4381 cagaacacag gaccggttct agagcgctgc caccatggac aagaagtaca gcatcggcct
     4441 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     4501 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     4561 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     4621 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     4681 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     4741 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     4801 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     4861 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     4921 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     4981 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     5041 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     5101 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     5161 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     5221 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     5281 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     5341 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     5401 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     5461 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     5521 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     5581 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     5641 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     5701 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     5761 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     5821 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     5881 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     5941 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     6001 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     6061 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     6121 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     6181 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     6241 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     6301 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     6361 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     6421 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     6481 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     6541 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     6601 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     6661 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     6721 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     6781 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     6841 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     6901 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     6961 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     7021 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     7081 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     7141 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     7201 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     7261 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     7321 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     7381 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     7441 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     7501 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     7561 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     7621 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     7681 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     7741 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     7801 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     7861 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     7921 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     7981 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     8041 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     8101 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     8161 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     8221 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     8281 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     8341 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     8401 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     8461 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     8521 gcgacctgcc gccacaaaga aggctggaca ggctaagaag aagaaagatt acaaagacga
     8581 tgacgataag ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga
     8641 agagaatcct ggaccgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     8701 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     8761 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     8821 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     8881 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     8941 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     9001 ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg
     9061 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     9121 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     9181 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     9241 gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     9301 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     9361 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     9421 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     9481 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     9541 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     9601 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     9661 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     9721 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     9781 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     9841 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     9901 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     9961 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
    10021 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
    10081 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
    10141 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
    10201 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
    10261 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
    10321 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
    10381 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
    10441 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    10501 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    10561 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    10621 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    10681 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    10741 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    10801 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    10861 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    10921 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10981 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    11041 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    11101 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    11161 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    11221 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    11281 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    11341 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    11401 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    11461 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    11521 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    11581 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    11641 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    11701 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    11761 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    11821 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    11881 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11941 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    12001 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    12061 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    12121 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    12181 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    12241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    12301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    12361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    12421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    12481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    12541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    12601 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12841 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caa
//
