LOCUS       sequence-440259-       14991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express fussed dCas9-TurboID for proximity labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14991)
  AUTHORS   Sun T, Xu Y, Xiang Y, Ou J, Soderblom EJ, Diao Y
  TITLE     Crosstalk between RNA m(6)A and DNA methylation regulates 
            transposable element chromatin activation and cell fate in human 
            pluripotent stem cells.
  JOURNAL   Nat Genet. 2023 Aug;55(8):1324-1335. doi: 
            10.1038/s41588-023-01452-5. Epub 2023 Jul 20.
  PUBMED    37474847
REFERENCE   2  (bases 1 to 14991)
  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: 
            "10.1038/s41588-023-01452-5"; journalName: "Nat Genet"; date: 
            "2023-08"; volume: "55"; issue: "8"; pages: "1324-1335"
COMMENT     Sequence Label: Lenti-dCas9-TurboID-blast
FEATURES             Location/Qualifiers
     source          1..14991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..87
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             103..156
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             157..555
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLNPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    580..1168
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(633..653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1051..1062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1171..1187)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1172..1188)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1693..1873
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1899..1916)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1905..2129
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2175..2603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2262..2281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2472..2493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2612..2632)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2617..2946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2797..2932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2859..2878
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2994..3041
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3060..3434
                     /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    3564..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3601..3620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3655..3674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3734..3750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3734..3750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3747..3769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3758..3774
                     /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(3782..3788,3789..3806,3807..3812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3827..3848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3965..3982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4136..4724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4216..4235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4895..5686,5687..5755))
                     /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(5756..5860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5935..5954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6126..6505
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6507..6705
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6660..6680
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6723..6903
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6950..7075
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7568..7801
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7986..8030
                     /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    8328..8445
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8575..9753
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8806..9744
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     9701..9721
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             9769..9816
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(9817..9840,9841..9843,9844..12399,12400..12402,
                     12403..13917)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             13972..13992
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14035..14991
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
ORIGIN
        1 gagagagaaa ttcaccaaga gacccatcct gattcagaga ctgcatttga aatcaaatca
       61 tcagttccga aaaagaaacg caaagtttgt acaggcagtg gagagggcag aggaagtctg
      121 ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg ccaagccttt gtctcaagaa
      181 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
      241 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
      301 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
      361 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
      421 ccctgcggac ggtgccgaca ggtgcttctc gatctgaatc ctgggatcaa agccatagtg
      481 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
      541 gtgtgggagg gctaagaatt cgatatcaag cttatcggta atcaacctct ggattacaaa
      601 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
      661 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
      721 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
      781 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
      841 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
      901 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
      961 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     1021 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     1081 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     1141 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgaga cctagaaaaa
     1201 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     1261 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     1321 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     1381 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     1441 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
     1501 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
     1561 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     1621 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
     1681 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     1741 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     1801 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     1861 aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     1921 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1981 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2041 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2101 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2161 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2221 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2281 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2341 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2401 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2461 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2521 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2581 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2881 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2941 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     3001 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     3061 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg
     3121 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg
     3181 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca
     3241 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg
     3301 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc
     3361 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg
     3421 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     3481 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3541 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3601 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3661 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     3721 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3781 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3841 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4561 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5881 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     5941 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     6001 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     6061 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     6121 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     6181 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6361 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6421 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     6541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6601 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6661 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
     6721 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6781 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6841 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6901 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     6961 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     7021 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     7081 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     7141 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     7201 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     7261 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     7321 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     7381 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     7441 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     7501 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     7561 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     7621 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     7681 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7741 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7801 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     7861 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     7921 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     7981 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     8041 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     8101 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     8161 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     8221 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     8281 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     8341 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     8401 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     8461 gacagcagag atccagtttg gttaattagc tagctgcaaa gatggataaa gttttaaaca
     8521 gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc
     8581 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     8641 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     8701 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     8761 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
     8821 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
     8881 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
     8941 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
     9001 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     9061 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     9121 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     9181 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     9241 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     9301 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     9361 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     9421 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     9481 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     9541 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     9601 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     9661 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     9721 caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgaa aaggccggcg
     9781 gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaca agaagtacag catcggcctg
     9841 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     9901 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     9961 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
    10021 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
    10081 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
    10141 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
    10201 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
    10261 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
    10321 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
    10381 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
    10441 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
    10501 cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc
    10561 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
    10621 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
    10681 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
    10741 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
    10801 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
    10861 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
    10921 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
    10981 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
    11041 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
    11101 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
    11161 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
    11221 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
    11281 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
    11341 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
    11401 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
    11461 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
    11521 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
    11581 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
    11641 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
    11701 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
    11761 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
    11821 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
    11881 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
    11941 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
    12001 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
    12061 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
    12121 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
    12181 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
    12241 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
    12301 atcaaccggc tgtccgacta cgatgtggac cacatcgtgc ctcagagctt tctgaaggac
    12361 gactccatcg acaacaaggt gctgaccaga agcgacaagg cccggggcaa gagcgacaac
    12421 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
    12481 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
    12541 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
    12601 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
    12661 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
    12721 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
    12781 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
    12841 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
    12901 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
    12961 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
    13021 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
    13081 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
    13141 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
    13201 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
    13261 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
    13321 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
    13381 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
    13441 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
    13501 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
    13561 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
    13621 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
    13681 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
    13741 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
    13801 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
    13861 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacagc
    13921 gctggaggag gtggaagcgg aggaggagga agcggaggag gaggtagcgg acctaagaaa
    13981 aagaggaagg tggcggccgc tggatccgat gctaagtcac tgactgcctg gtccaaagac
    14041 aatactgtgc ctctgaagct gatcgctctc ctggctaatg gcgagttcca tagtggcgaa
    14101 cagctgggag aaaccctggg catgtccagg gccgctatca acaagcacat tcagactctg
    14161 cgcgactggg gcgtggacgt gttcaccgtg cccggaaagg gctactctct gcccgagcct
    14221 atcccgctgc tgaacgctaa acagattctg ggacagctgg acggcgggag cgtggcagtc
    14281 ctgcctgtgg tcgactccac caatcagtac ctgctggatc gaatcggcga gctgaagagt
    14341 ggggatgctt gcattgcaga atatcagcag gcagggagag gaagcagagg gaggaaatgg
    14401 ttctctcctt ttggagctaa cctgtacctg agtatgtttt ggcgcctgaa gcggggacca
    14461 gcagcaatcg gcctgggccc ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg
    14521 ggagcagaca aggtgcgagt caaatggccc aatgacctgt atctgcagga tagaaagctg
    14581 gcaggcatcc tggtggagct ggccggaata acaggcgatg ctgcacagat cgtcattggc
    14641 gccgggatta acgtggctat gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc
    14701 acactgcagg aagcagggat taacctggac aggaatactc tggccgctac gctgatccga
    14761 gagctgcggg cagccctgga actgttcgag caggaaggcc tggctccata tctgccacgg
    14821 tgggagaagc tggataactt catcaataga cccgtgaagc tgatcattgg ggacaaagag
    14881 attttcggga ttagccgggg gattgataaa cagggagccc tgctgctgga acaggacgga
    14941 gttatcaaac cctggatggg cggagaaatc agtctgcggt ctgccgaaaa g
//
