LOCUS       sequence-489439-       12816 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Knockout of human FAM136A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12816)
  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 12816)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 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_sg2
FEATURES             Location/Qualifiers
     source          1..12816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(61..105)
                     /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    290..523
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1016..1141
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1188..1368
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1386..1584)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1411..1431)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1586..1965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2141..2160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2235..2339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2340..2408,2409..3200)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2558..2577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3371..3959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3860..3879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4113..4130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4247..4268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4283..4288,4289..4306,4307..4313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4321..4337
                     /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)."
     primer_bind     4326..4348
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4345..4361
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4345..4361
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4410..4531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4421..4440)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4475..4494
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4661..5035)
                     /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"
     promoter        complement(5054..5101)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5149..5478)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5163..5298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5217..5236)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5463..5483
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5492..5920)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5602..5623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5814..5833
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5966..6190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6179..6196
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6222..6455
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    6527..7115
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6633..6644
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7042..7062
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7131..7526)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(7527..7583)
                     /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             complement(7593..7616)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(7617..7664)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(7665..11768)
                     /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             complement(7665..11768)
                     /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"
     regulatory      11765..11774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(11793..12004)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(12066..12141)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(12163..12411)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(12221..12240)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(12391..12411)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    12462..12579
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
       61 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
      121 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
      181 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
      241 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
      301 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
      361 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
      421 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
      481 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
      541 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
      601 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg
      661 tctgaagatc agcggccgct tgctgtgcgg tggtcttact tttgttttgc tcttcctcta
      721 tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg
      781 ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta
      841 gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga
      901 atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc
      961 ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc
     1021 tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca
     1081 gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga
     1141 gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt
     1201 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
     1261 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
     1321 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa
     1381 acgcgctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     1441 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     1501 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     1561 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     1621 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     1681 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     1741 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     1801 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     1861 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     1921 accccgtaat tgattactat taataactag tcaataatca atgtcaacgc gcgcgtatat
     1981 ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa
     2041 ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc
     2101 tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta
     2161 ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt
     2221 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2281 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2341 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2401 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2461 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2521 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2581 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2641 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2701 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2761 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2821 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2881 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2941 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3001 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3061 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3121 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3181 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3241 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3301 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3361 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3421 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3481 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3541 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3601 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3661 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3721 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3781 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3841 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3901 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3961 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4021 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4081 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4141 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4201 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     4261 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     4321 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     4381 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
     4441 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4501 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
     4561 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     4621 catagaaggc ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca
     4681 cgaagtgcac gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct
     4741 cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg
     4801 accactcggc gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg
     4861 gcaccacctg gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg
     4921 cgaagtcgtc ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg
     4981 ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt
     5041 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     5101 cacgtgctga tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc
     5161 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc
     5221 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5281 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5341 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5401 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5461 acaccctaac tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc
     5521 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5581 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5641 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5701 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5761 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5821 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5881 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc
     5941 cagctggttc tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta
     6001 ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
     6061 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
     6121 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
     6181 gaaggcacag tcgaggctga tcagcgggtt taaacgggcc ctgctagaga ttttccacac
     6241 tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac
     6301 tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc
     6361 agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacaag caaaaagcag
     6421 atcttgtctt cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg
     6481 gctaagatct acagctgcct tgtaagtcat tggtcttaaa gtcgacgcgg ggaggcggcc
     6541 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
     6601 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
     6661 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat
     6721 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
     6781 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
     6841 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
     6901 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
     6961 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     7021 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     7081 aatctttcac aaattttgta atccagaggt tgattgtcga cttaacgcgt ttagccctcc
     7141 cacacataac cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg
     7201 tccttcacta tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg
     7261 caggggctca agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca
     7321 gctgccgcag cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa
     7381 tgatatacat tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg
     7441 ctgtagtctt cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg
     7501 gattcttctt gagacaaagg cttggccggt ccaggattct cttcgacatc tccggcttgt
     7561 ttcagcagag agaagtttgt tgcgccggat cccttatcgt catcgtcttt gtaatctttc
     7621 ttcttcttag cctgtccagc cttctttgtg gcggcaggtc gcttgtcgcc tcccagctga
     7681 gacaggtcga tccgtgtctc gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc
     7741 agcacctctt tggtgctggt gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag
     7801 gcggcagggg ctcccagatt ggtcagggta aacaggtgga tgatattctc ggcctgctct
     7861 ctgatgggct tatcccggtg cttgttgtag gcggacagca ctttgtccag attagcgtcg
     7921 gccaggatca ctctcttgga gaactcgctg atctgctcga tgatctcgtc caggtagtgc
     7981 ttgtgctgtt ccacaaacag ctgtttctgc tcattatcct cgggggagcc cttcagcttc
     8041 tcatagtggc tggccaggta caggaagttc acatatttgg agggcagggc cagttcgttt
     8101 cccttctgca gttcgccggc agaggccagc attctcttcc ggccgttttc cagctcgaac
     8161 agggagtact taggcagctt gatgatcagg tcctttttca cttctttgta gcccttggct
     8221 tccagaaagt cgatgggatt cttctcgaag ctgcttcttt ccatgatggt gatccccagc
     8281 agctctttca cactcttcag tttcttggac ttgccctttt ccactttggc caccaccagc
     8341 acagaatagg ccacggtggg gctgtcgaag ccgccgtact tcttagggtc ccagtccttc
     8401 tttctggcga tcagcttatc gctgttcctc ttgggcagga tagactcttt gctgaagccg
     8461 cctgtctgca cctcggtctt tttcacgata ttcacttggg gcatgctcag cactttccgc
     8521 acggtggcaa aatcccggcc cttatcccac acgatctccc cggtttcgcc gtttgtctcg
     8581 atcagaggcc gcttccggat ctcgccgttg gccagggtaa tctcggtctt gaaaaagttc
     8641 atgatgttgc tgtagaagaa gtacttggcg gtagccttgc cgatttcctg ctcgctcttg
     8701 gcgatcatct tccgcacgtc gtacaccttg tagtcgccgt acacgaactc gctttccagc
     8761 ttagggtact ttttgatcag ggcggttccc acgacggcgt tcaggtaggc gtcgtgggcg
     8821 tggtggtagt tgttgatctc gcgcactttg taaaactgga aatccttccg gaaatcggac
     8881 accagcttgg acttcagggt gatcactttc acttcccgga tcagcttgtc attctcgtcg
     8941 tacttagtgt tcatccggga gtccaggatc tgtgccacgt gctttgtgat ctgccgggtt
     9001 tccaccagct gtctcttgat gaagccggcc ttatccagtt cgctcaggcc gcctctctcg
     9061 gccttggtca gattgtcgaa ctttctctgg gtaatcagct tggcgttcag cagctgccgc
     9121 cagtagttct tcatcttctt cacgacctct tcggagggca cgttgtcgct cttgccccgg
     9181 ttcttgtcgc ttctggtcag caccttgttg tcgatggagt cgtccttcag aaagctctga
     9241 ggcacgatat ggtccacatc gtagtcggac agccggttga tgtccagttc ctggtccacg
     9301 tacatatccc gcccattctg caggtagtac aggtacagct tctcgttctg cagctgggtg
     9361 ttttccacgg ggtgttcttt caggatctgg ctgcccagct ctttgatgcc ctcttcgatc
     9421 cgcttcattc tctcgcggct gttcttctgt cccttctggg tggtctggtt ctctctggcc
     9481 atttcgatca cgatgttctc gggcttgtgc cggcccatca ctttcacgag ctcgtccacc
     9541 accttcactg tctgcaggat gcccttctta atggcggggc tgccggccag attggcaatg
     9601 tgctcgtgca ggctatcgcc ctggccggac acctgggctt tctggatgtc ctctttaaag
     9661 gtcaggctgt cgtcgtggat cagctgcatg aagtttctgt tggcgaagcc gtcggacttc
     9721 aggaaatcca ggattgtctt gccggactgc ttgtcccgga tgccgttgat cagcttccgg
     9781 ctcagcctgc cccagccggt gtatctccgc cgcttcagct gcttcatcac tttgtcgtcg
     9841 aacaggtggg cataggtttt cagccgttcc tcgatcatct ctctgtcctc aaacagtgtc
     9901 agggtcagca cgatatcttc cagaatgtcc tcgttttcct cattgtccag gaagtccttg
     9961 tccttgataa ttttcagcag atcgtggtat gtgcccaggg aggcgttgaa ccgatcttcc
    10021 acgccggaga tttccacgga gtcgaagcac tcgattttct tgaagtagtc ctctttcagc
    10081 tgcttcacgg tcactttccg gttggtcttg aacagcaggt ccacgatggc ctttttctgc
    10141 tcgccgctca ggaaggcggg ctttctcatt ccctcggtca cgtatttcac tttggtcagc
    10201 tcgttataca cggtgaagta ctcgtacagc aggctgtgct tgggcagcac cttctcgttg
    10261 ggcaggttct tatcgaagtt ggtcatccgc tcgatgaagc tctgggcgga agcgcccttg
    10321 tccaccactt cctcgaagtt ccagggggtg atggtttcct cgctctttct ggtcatccag
    10381 gcgaatctgc tgtttcccct ggccagaggg cccacgtagt aggggatgcg gaaggtcagg
    10441 atcttctcga tcttttcccg gttgtccttc aggaatgggt aaaaatcttc ctgccgccgc
    10501 agaatggcgt gcagctctcc caggtggatc tggtggggga tgctgccgtt gtcgaaggtc
    10561 cgctgcttcc gcagcaggtc ctctctgttc agcttcacga gcagttcctc ggtgccgtcc
    10621 atcttttcca ggatgggctt gatgaacttg tagaactctt cctggctggc tccgccgtca
    10681 atgtagccgg cgtagccgtt cttgctctgg tcgaagaaaa tctctttgta cttctcaggc
    10741 agctgctgcc gcacgagagc tttcagcagg gtcaggtcct ggtggtgctc gtcgtatctc
    10801 ttgatcatag aggcgctcag gggggccttg gtgatctcgg tgttcactct caggatgtcg
    10861 ctcagcagga tggcgtcgga caggttcttg gcggccagaa acaggtcggc gtactggtcg
    10921 ccgatctggg ccagcaggtt gtccaggtcg tcgtcgtagg tgtccttgct cagctgcagt
    10981 ttggcatcct cggccaggtc gaagttgctc ttgaagttgg gggtcaggcc caggctcagg
    11041 gcaatcaggt ttccgaacag gccattcttc ttctcgccgg gcagctgggc gatcagattt
    11101 tccagccgtc tgctcttgct cagtctggca gacaggatgg ccttggcgtc cacgccgctg
    11161 gcgttgatgg ggttttcctc gaacagctgg ttgtaggtct gcaccagctg gatgaacagc
    11221 ttgtccacgt cgctgttgtc ggggttcagg tcgccctcga tcaggaagtg gccccggaac
    11281 ttgatcatgt gggccagggc cagatagatc agccgcaggt cggccttgtc ggtgctgtcc
    11341 accagtttct ttctcaggtg gtagatggtg gggtacttct cgtggtaggc cacctcgtcc
    11401 acgatgttgc cgaagatggg gtgccgctcg tgcttcttat cctcttccac caggaaggac
    11461 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
    11521 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
    11581 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
    11641 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
    11701 tcggtgatca cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc
    11761 ttgtccatgg tggcagcgct ctagaaccgg tcctgtgttc tggcggcaaa cccgttgcga
    11821 aaaagaacgt tcacggcgac tactgcactt atatacggtt ctcccccacc ctcgggaaaa
    11881 aggcggagcc agtacacgac atcactttcc cagtttaccc cgcgccacct tctctaggca
    11941 ccggatcaat tgccgacccc tccccccaac ttctcgggga ctgtgggcga tgtgcgctct
    12001 gcccactgac gggcaccgga gccaattccc actcctttca agacctagct agcgaattca
    12061 aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct tattttaact
    12121 tgctatttct agctctaaaa cccatgaagc aggtgcacca gcggtgtttc gtcctttcca
    12181 caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc
    12241 cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact ttctacgtca
    12301 cgtattttgt actaatatct ttgtgtttac agtcaaatta attctaatta tctctctaac
    12361 agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct cggtacctta
    12421 attaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta
    12481 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
    12541 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
    12601 tgtctgcaga attggcgcac gcagtgccga tccgttcact aatcgaatgg atctgtctct
    12661 gtctctctct ccaccttctt cttctattcc ttcgggcctg tcgggtcccc tcggggttgg
    12721 gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag
    12781 tacagcaaaa actattctta aacctaccaa gcctcc
//
