LOCUS       sequence-459920-       13763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for dCas9 fused to miniTurboNLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13763)
  AUTHORS   Cenik BK, Aoi Y, Iwanaszko M, Howard BC, Morgan MA, Andersen GD, 
            Bartom ET, Shilatifard A
  TITLE     TurboCas: A method for locus-specific labeling of genomic regions 
            and isolating their associated protein interactome.
  JOURNAL   Mol Cell. 2024 Dec 19;84(24):4929-4944.e8. doi: 
            10.1016/j.molcel.2024.11.007.
  PUBMED    39706164
REFERENCE   2  (bases 1 to 13763)
  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.molcel.2024.11"; journalName: "Mol Cell"; date: 
            "2024-12-19- 19"; volume: "84"; issue: "24"; pages: "4929-4944.e8"
COMMENT     Sequence Label: Lenti-TurboCas-NLS-Blast
FEATURES             Location/Qualifiers
     source          1..13763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             119..299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(325..342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    331..555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      601..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(688..707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     898..919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1038..1058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1203..1337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1264..1283
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1399..1446
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1465..1839
                     /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    1969..2090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2006..2025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2060..2079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2139..2155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2139..2155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2152..2174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2163..2179
                     /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(2187..2193,2194..2211,2212..2217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2232..2253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2621..2640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3300..4091,4092..4160))
                     /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"
     promoter        complement(4161..4265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4340..4359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4531..4910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4912..5110
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5065..5085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5128..5308
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5355..5480
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5973..6206
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6391..6435
                     /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    6733..6850
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6943..7154
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7183..7209
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7213..7239
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7243..7269
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7276..8046
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             8053..8094
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(8095..8121,8122..8124,8125..10608,10609..10614,
                     10615..10680,10681..10683,10684..12198)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLPKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8095..12198
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLPKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             12205..12225
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12229..12249
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12253..12273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12289..12345
                     /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             12346..12744
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    12769..13357
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12822..12842)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13240..13251)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13360..13376)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13361..13377)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
       61 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
      121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      301 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
      361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
      421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
      481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
      541 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
      601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      961 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1201 agtcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1261 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     1321 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     1381 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     1441 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     1501 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     1561 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     1621 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     1681 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     1741 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     1801 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     1861 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1921 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1981 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2101 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2161 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2221 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2281 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4321 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4381 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4441 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4501 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4561 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4681 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4741 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4801 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4861 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4921 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4981 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5041 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5101 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     5161 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5221 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5281 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     5341 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     5401 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     5461 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     5521 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     5581 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     5641 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     5701 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     5761 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     5821 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     5881 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     5941 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     6001 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     6061 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     6121 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     6181 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     6241 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     6301 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     6361 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     6421 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     6481 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     6541 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     6601 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     6661 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
     6721 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     6781 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     6841 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttagctagct
     6901 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
     6961 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
     7021 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     7081 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     7141 ccgccagaac acaggaccgg ttctagagcg gccgccacca tgtacccgta tgatgttccg
     7201 gattacgctg gctatcccta cgacgtgccc gactatgccg ggtaccccta tgacgtccca
     7261 gactacgcag ctagcatccc gctgctgaac gctaaacaga ttctgggaca gctggacggc
     7321 gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc agtacctgct ggatcgaatc
     7381 ggcgagctga agagtgggga tgcttgcatt gcagaatatc agcaggcagg gagaggaagc
     7441 agagggagga aatggttctc tccttttgga gctaacctgt acctgagtat gttttggcgc
     7501 ctgaagcggg gaccagcagc aatcggcctg ggcccggtca tcggaattgt catggcagaa
     7561 gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat ggcccaatga cctgtatctg
     7621 caggatagaa agctggcagg catcctggtg gagctggccg gaataacagg cgatgctgca
     7681 cagatcgtca ttggcgccgg gattaacgtg gctatgaggc gcgtggagga aagcgtggtc
     7741 aatcagggct ggatcacact gcaggaagca gggattaacc tggacaggaa tactctggcc
     7801 gctatgctga tccgagagct gcgggcagcc ctggaactgt tcgagcagga aggcctggct
     7861 ccatatctgt cacggtggga gaagctggat aacttcatca atagacccgt gaagctgatc
     7921 attggggaca aagagatttt cgggattagc cgggggattg ataaacaggg agccctgctg
     7981 ctggaacagg acggagttat caaaccctgg atgggcggag aaatcagtct gcggtctgcc
     8041 gaaaaggaat tcggcaagcc catccccaac cccctgctgg gcctggacag caccatggac
     8101 aagaagtact ccattgggct cgccatcggc acaaacagcg tcggctgggc cgtcattacg
     8161 gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc
     8221 ataaagaaga acctcattgg cgccctcctg ttcgactccg gggaaaccgc cgaagccacg
     8281 cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg
     8341 caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag
     8401 gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc
     8461 gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt
     8521 gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc
     8581 aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac
     8641 aaactcttta tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac
     8701 gcatccggag ttgacgccaa agcaatcctg agcgctaggc tgcccaaatc ccggcggctc
     8761 gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc
     8821 gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc
     8881 aagcttcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc
     8941 ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg
     9001 agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc
     9061 aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa
     9121 ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac
     9181 attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa
     9241 atggacggca ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag
     9301 cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc
     9361 ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa
     9421 atcctcacat ttcggatacc ctactatgta ggccccctcg cccggggaaa ttccagattc
     9481 gcgtggatga ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg
     9541 gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg
     9601 cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac
     9661 gagctcacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga
     9721 gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa
     9781 cagctcaaag aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga
     9841 gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa
     9901 gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc
     9961 cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc
    10021 ttcgacgaca aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg
    10081 tcaagaaaac tgatcaatgg gatccgagac aagcagagtg gaaagacaat cctggatttt
    10141 cttaagtccg atggatttgc caaccggaac ttcatgcagt tgatccatga tgactctctc
    10201 acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag
    10261 cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag
    10321 gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag
    10381 atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag
    10441 aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa
    10501 aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg
    10561 tacgtggatc aggaactgga catcaatcgg ctctccgact acgacgtgga tgccatcgtg
    10621 ccccagtctt ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa
    10681 aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat
    10741 tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag
    10801 gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt
    10861 gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag
    10921 tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg
    10981 gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac
    11041 catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc
    11101 aagcttgaat ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc
    11161 gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt
    11221 atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt
    11281 atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca
    11341 gtccggaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc
    11401 ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc
    11461 aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt
    11521 gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa
    11581 ctgctgggca tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc
    11641 gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct
    11701 ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa
    11761 ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat
    11821 gaaaagctca aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac
    11881 aaacactacc ttgatgagat catcgagcaa ataagcgaat tctccaaaag agtgatcctc
    11941 gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc
    12001 agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca
    12061 gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc
    12121 ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc
    12181 tctcagctcg gtggagactg tacacccaag aagaagagga aggtggaccc caagaagaag
    12241 aggaaggtgg accccaagaa gaagaggaag gtgtgtacag gatccggcgc aacaaacttc
    12301 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatggc caagcctttg
    12361 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
    12421 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
    12481 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
    12541 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
    12601 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
    12661 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
    12721 tctggttatg tgtgggaggg ctaagaattc gatatcaagc ttatcggtaa tcaacctctg
    12781 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
    12841 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
    12901 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
    12961 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
    13021 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
    13081 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
    13141 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
    13201 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
    13261 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
    13321 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgagac
    13381 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
    13441 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
    13501 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
    13561 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
    13621 acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca
    13681 ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc
    13741 aatgaaggag agaacacccg ctt
//
