LOCUS       sequence-489423-       12815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout of human FAM136A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12815)
  AUTHORS   Harhai M, Foged MM, Zarges C, Landoni JC, Chollet S, Simonelli M, 
            Recazens E, Lisci M, Laban N, Manley S, Riemer J, Lopez-Escamez JA, 
            Lysakowski A, Jourdain AA
  TITLE     An updated inventory of genes essential for oxidative 
            phosphorylation identifies a mitochondrial origin in familial 
            Meniere's disease.
  JOURNAL   Cell Rep. 2025 Jul 25;44(8):116069. doi: 
            10.1016/j.celrep.2025.116069.
  PUBMED    40714634
REFERENCE   2  (bases 1 to 12815)
  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: "Cell Rep.";
            date: "2025-07-25"; volume: "44(8):116069. doi"; pages: " 
            10.1016/j.celrep.2025.116069"
COMMENT     Sequence Label: pLentiCRISPR_blast_FAM136A_sg1
FEATURES             Location/Qualifiers
     source          1..12815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..68
                     /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             69..464
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    480..1068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(533..553)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(951..962)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1140..1373
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1399..1416)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1405..1629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1675..2103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1762..1781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1972..1993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2112..2132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2117..2446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2297..2432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2359..2378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2494..2541
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2560..2934
                     /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    3064..3185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3155..3174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3234..3250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3234..3250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3247..3269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3258..3274
                     /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(3282..3288,3289..3306,3307..3312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3327..3348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3465..3482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3636..4224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3716..3735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4395..5186,5187..5255))
                     /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     5018..5037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5256..5360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5435..5454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5630..6009
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6011..6209
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6164..6184
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6227..6407
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6454..6579
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7072..7305
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7490..7534
                     /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    7832..7949
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8000..8248
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8000..8020
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8171..8190
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8269..8344
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8406..8617
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8636..8645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8642..12745
                     /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             8642..12745
                     /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             12746..12793
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(12794..12815,1..2)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 agggatccgg cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc gaagagaatc
       61 ctggaccggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
      121 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
      181 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
      241 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
      301 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
      361 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
      421 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaaacgcgt taagtcgaca
      481 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
      541 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
      601 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
      661 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
      721 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
      781 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
      841 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
      901 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
      961 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     1021 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     1081 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     1141 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     1201 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     1261 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     1321 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg
     1381 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1441 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1501 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1561 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     1621 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     1681 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1741 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1801 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1861 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1921 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1981 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2041 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2101 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2161 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2281 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2341 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2401 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     2461 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     2521 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     2581 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     2641 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     2701 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     2761 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     2821 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     2881 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     2941 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3001 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3061 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3121 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3181 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3241 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3301 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3361 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3421 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3481 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3541 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4441 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4501 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4561 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4621 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4681 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4741 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4801 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4861 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4921 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4981 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5041 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5101 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5161 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5221 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5281 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5341 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     5401 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5461 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     5521 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     5581 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgcgcgttg acattgatta
     5641 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5701 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5761 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5821 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     5881 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     5941 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6001 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     6061 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     6121 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     6181 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
     6241 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6301 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6361 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     6421 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     6481 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     6541 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     6601 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     6661 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     6721 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     6781 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     6841 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     6901 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     6961 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     7021 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     7081 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     7141 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     7201 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     7261 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     7321 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     7381 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     7441 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     7501 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     7561 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     7621 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     7681 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     7741 agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat
     7801 tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     7861 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     7921 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     7981 tttggttaat taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     8041 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     8101 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     8161 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     8221 atatatcttg tggaaaggac gaaacaccgc atggtgcacc gggccagggt tttagagcta
     8281 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     8341 gtgctttttt gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     8401 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     8461 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     8521 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     8581 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac
     8641 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     8701 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     8761 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     8821 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     8881 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     8941 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     9001 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     9061 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     9121 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     9181 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     9241 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     9301 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     9361 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     9421 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     9481 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     9541 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     9601 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     9661 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     9721 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     9781 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     9841 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     9901 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     9961 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
    10021 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
    10081 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
    10141 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
    10201 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
    10261 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
    10321 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
    10381 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
    10441 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
    10501 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
    10561 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
    10621 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
    10681 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
    10741 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
    10801 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
    10861 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
    10921 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
    10981 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
    11041 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
    11101 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
    11161 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
    11221 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
    11281 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
    11341 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
    11401 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
    11461 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
    11521 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
    11581 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
    11641 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
    11701 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
    11761 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
    11821 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
    11881 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
    11941 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
    12001 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
    12061 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
    12121 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
    12181 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
    12241 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
    12301 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
    12361 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
    12421 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
    12481 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
    12541 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
    12601 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
    12661 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
    12721 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
    12781 taagaagaag aaagattaca aagacgatga cgata
//
