LOCUS       sequence-373292-       15024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 effector fusion for manipulating transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15024)
  AUTHORS   Tycko J, Van MV, Aradhana, DelRosso N, Ye H, Yao D, Valbuena R, 
            Vaughan-Jackson A, Xu X, Ludwig C, Spees K, Liu K, Gu M, Khare V, 
            Mukund AX, Suzuki PH, Arana S, Zhang C, Du PP, Ornstein TS, Hess GT,
            Kamber RA, Qi LS, Khalil AS, Bintu L, Bassik MC
  TITLE     Development of compact transcriptional effectors using 
            high-throughput measurements in diverse contexts.
  JOURNAL   Nat Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6.
  PUBMED    39487265
REFERENCE   2  (bases 1 to 15024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6."
COMMENT     Sequence Label: pJT337 Lenti pEF-dCas9-3XFLAG-ZN471
            KRAB-T2A-t...
FEATURES             Location/Qualifiers
     source          1..15024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        158..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     692..712
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             755..935
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    982..1107
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1600..1833
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2018..2062
                     /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    2360..2477
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2607..3785
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2838..3776
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3733..3753
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3801..3848
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3849..3872,3873..3875,3876..6431,6432..6434,
                     6435..7949)
                     /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             8004..8024
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8040..8105
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8376..8429
                     /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             9156..9212
                     /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             9213..9611
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9636..10224
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9689..9709)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10107..10118)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10227..10243)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10228..10244)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10749..10929
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10955..10972)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10961..11185
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11231..11659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11318..11337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11528..11549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11668..11688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11673..12002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11853..11988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11915..11934
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12050..12097
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12116..12490
                     /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    12620..12741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12657..12676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12711..12730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12790..12806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12790..12806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12803..12825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12814..12830
                     /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(12838..12844,12845..12862,12863..12868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12883..12904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13021..13038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13192..13780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13272..13291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13951..14742,14743..14811))
                     /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     14574..14593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14812..14916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14991..15010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
       61 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
      121 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      541 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      601 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      661 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      721 gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg
      781 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      841 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      901 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      961 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     1021 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     1081 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     1141 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     1201 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1261 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1321 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1381 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1441 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1501 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1561 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1621 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1681 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1741 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1801 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1861 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1921 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1981 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     2041 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     2101 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     2161 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     2221 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     2281 agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa
     2341 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     2401 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     2461 caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta
     2521 gctagctgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     2581 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     2641 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     2701 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2761 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2821 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     2881 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     2941 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     3001 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     3061 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     3121 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     3181 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     3241 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     3301 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     3361 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     3421 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     3481 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     3541 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     3601 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     3661 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     3721 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     3781 cgtgacgtac ggccaccatg aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
     3841 agaaaaagga caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg
     3901 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     3961 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     4021 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     4081 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     4141 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     4201 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     4261 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     4321 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     4381 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     4441 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     4501 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     4561 gcaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     4621 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     4681 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     4741 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     4801 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     4861 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     4921 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     4981 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     5041 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     5101 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     5161 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     5221 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     5281 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     5341 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     5401 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     5461 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     5521 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     5581 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     5641 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     5701 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     5761 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     5821 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     5881 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     5941 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     6001 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     6061 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     6121 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     6181 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     6241 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     6301 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     6361 accacatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     6421 gaagcgacaa ggcccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     6481 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     6541 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     6601 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     6661 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     6721 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     6781 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     6841 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     6901 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     6961 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     7021 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     7081 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     7141 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     7201 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     7261 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     7321 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     7381 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     7441 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     7501 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     7561 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     7621 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     7681 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     7741 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     7801 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     7861 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     7921 ggatcgacct gtctcagctg ggaggcgaca gcgctggagg aggtggaagc ggaggaggag
     7981 gaagcggagg aggaggtagc ggacctaaga aaaagaggaa ggtggcggcc gctggatccg
     8041 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     8101 acaagggtgg atccggatca ggctccaacg tggaggtggt gaaggtgatg ccccaggacc
     8161 tggtgacctt caaggacgtg gccatcgact tcagccagga ggagtggcag tggatgaacc
     8221 ccgcccagaa gagactgtac agaagcatga tgctggagaa ctaccagagc ctggtgagcc
     8281 tgggcctgtg catcagcaag ccctacgtga tcagcctgct ggagcagggc agagagccct
     8341 gggagatgac cagcgagatg accagaggat ctggagaggg aaggggaagc ctcctaactt
     8401 gcggagatgt cgaggagaat cctggcccaa tggttagtaa gggagaggag ttgataaaag
     8461 agaatatgca catgaaactg tatatggaag ggacagtgga caaccatcac ttcaagtgca
     8521 cctctgaggg agaaggaaag ccatacgaag gcacccagac tatgcgaatt aaggtcgtgg
     8581 agggagggcc tctccccttc gccttcgata ttcttgcaac atctttcctc tacgggagca
     8641 aaaccttcat caatcacaca caaggcatac ctgacttttt taagcagagt tttccagagg
     8701 gctttacctg ggagcgtgtg actacatacg aagatggggg tgtgctgacg gctacacagg
     8761 atacttcact tcaggacggt tgtctgatct acaacgtcaa gatcaggggg gtcaatttca
     8821 cctcgaatgg accagtaatg caaaaaaaga ccctgggatg ggaagctttt actgaaactc
     8881 tctatcccgc cgacggcggc ctcgaaggca gaaacgatat ggctttgaag ctggttggcg
     8941 ggtcccatct aattgccaat gccaaaacca catataggag caaaaagcct gcaaagaacc
     9001 tgaagatgcc gggtgtgtac tatgttgact atagattaga gcggatcaaa gaggcaaata
     9061 acgaaacgta cgtggagcag cacgaggtag ccgtggcgcg ctactgcgat ttaccctcca
     9121 aactgggcca taaactgaac tgtacaggaa gcggagctac taacttcagc ctgctgaagc
     9181 aggctggcga cgtggaagaa aacccaggac ctatggccaa gcctttgtct caagaagaat
     9241 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     9301 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     9361 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     9421 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     9481 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     9541 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     9601 gggagggcta agaattcgat atcaagctta tcggtaatca acctctggat tacaaaattt
     9661 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     9721 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     9781 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     9841 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     9901 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     9961 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
    10021 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
    10081 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
    10141 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
    10201 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg
    10261 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
    10321 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
    10381 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
    10441 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
    10501 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
    10561 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
    10621 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
    10681 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
    10741 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
    10801 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
    10861 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
    10921 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
    10981 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
    11041 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
    11101 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
    11161 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
    11221 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
    11281 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
    11341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
    11401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
    11461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
    11521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
    11581 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
    11641 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
    11701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
    11761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
    11821 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
    11881 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
    11941 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
    12001 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
    12061 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
    12121 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
    12181 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
    12241 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
    12301 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
    12361 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
    12421 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
    12481 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
    12541 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
    12601 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
    12661 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    12721 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    12781 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    12841 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    12901 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    12961 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
    13021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    13081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    13141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    13201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    13261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    13321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    13381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    13441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    13501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    13561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    13621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    13681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    13741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    13801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    13861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    13921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    13981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    14041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    14101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    14161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    14221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    14281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    14341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    14401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    14461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    14521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    14581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    14641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    14701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    14761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    14821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    14881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    14941 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
    15001 tctgatgccg catagttaag ccag
//
