LOCUS       sequence-373289-       15219 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 15219)
  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 15219)
  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: pJT282 Lenti
            pEF-dCas9-3XFLAG-N+Z+F-T2A-tagBFP...
FEATURES             Location/Qualifiers
     source          1..15219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        625..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     778..798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             841..1021
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1068..1193
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2693..3871
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2924..3862
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3819..3839
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3887..3934
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3935..3958,3959..3961,3962..6517,6518..6520,
                     6521..8035)
                     /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             8090..8110
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8126..8191
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8657..8710
                     /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             9437..9493
                     /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             9494..9892
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9917..10505
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9970..9990)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10388..10399)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10508..10524)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10509..10525)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11030..11210
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11236..11253)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11242..11466
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11512..11940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11599..11618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11809..11830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11949..11969)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11954..12283
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12134..12269
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12196..12215
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12331..12378
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12397..12771
                     /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    12901..13022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12938..12957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12992..13011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13071..13087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13071..13087)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13084..13106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13095..13111
                     /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(13119..13125,13126..13143,13144..13149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13164..13185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13302..13319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13473..14061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13553..13572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14232..15023,15024..15092))
                     /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     14855..14874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15093..15197)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
      841 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
      901 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      961 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     1021 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     1081 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     1141 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     1201 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     1261 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     1321 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     1381 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     1441 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     1501 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt taattagcta gctgcaaaga tggataaagt tttaaacaga
     2641 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
     2701 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     2761 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     2821 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     2881 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     2941 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     3001 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     3061 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     3121 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     3181 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     3241 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     3301 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     3361 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     3421 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     3481 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     3541 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     3601 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     3661 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     3721 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     3781 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     3841 aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatgaaaa ggccggcggc
     3901 cacgaaaaag gccggccagg caaaaaagaa aaaggacaag aagtacagca tcggcctggc
     3961 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     4021 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     4081 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     4141 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     4201 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     4261 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     4321 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     4381 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     4441 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     4501 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     4561 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     4621 cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc
     4681 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     4741 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     4801 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     4861 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     4921 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     4981 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     5041 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     5101 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     5161 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     5221 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     5281 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     5341 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     5401 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     5461 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     5521 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     5581 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     5641 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     5701 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     5761 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     5821 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     5881 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     5941 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     6001 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     6061 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     6121 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     6181 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     6241 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     6301 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     6361 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     6421 caaccggctg tccgactacg atgtggacca catcgtgcct cagagctttc tgaaggacga
     6481 ctccatcgac aacaaggtgc tgaccagaag cgacaaggcc cggggcaaga gcgacaacgt
     6541 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     6601 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     6661 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     6721 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     6781 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     6841 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     6901 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     6961 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     7021 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     7081 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     7141 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     7201 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     7261 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     7321 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     7381 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     7441 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     7501 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     7561 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     7621 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     7681 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     7741 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     7801 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     7861 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     7921 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     7981 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacagcgc
     8041 tggaggaggt ggaagcggag gaggaggaag cggaggagga ggtagcggac ctaagaaaaa
     8101 gaggaaggtg gcggccgctg gatccgacta caaagaccat gacggtgatt ataaagatca
     8161 tgacatcgac tacaaggatg acgatgacaa gggtggatcc ggatcaggct ccgagggcca
     8221 gagcgacgag agagccctgc tggaccagct gcacaccctg ctgagcaaca ccgacgccac
     8281 cggcctggag gagatcgaca gagccctggg catccccgag ctggtgaacc agggccaggc
     8341 cctggagccc aagcagggag gaggaggtag cggattcgtg accctgaagg acgtgggcat
     8401 ggacttcacc ctgggcgact gggagcagct gggcctggag cagggcgaca ccttctggga
     8461 caccgccctg gacaactgcc aggacctgtt cctgctggac ccccccggag gaggaggtag
     8521 cggacacgag aagttcccca gcgacctgga cctggacatg ttcaacggca gcctggagtg
     8581 cgacatggag agcatcatca gaagcgagct gatggacgcc gacggcctgg acttcaactt
     8641 cgacagcgga tctggagagg gaaggggaag cctcctaact tgcggagatg tcgaggagaa
     8701 tcctggccca atggttagta agggagagga gttgataaaa gagaatatgc acatgaaact
     8761 gtatatggaa gggacagtgg acaaccatca cttcaagtgc acctctgagg gagaaggaaa
     8821 gccatacgaa ggcacccaga ctatgcgaat taaggtcgtg gagggagggc ctctcccctt
     8881 cgccttcgat attcttgcaa catctttcct ctacgggagc aaaaccttca tcaatcacac
     8941 acaaggcata cctgactttt ttaagcagag ttttccagag ggctttacct gggagcgtgt
     9001 gactacatac gaagatgggg gtgtgctgac ggctacacag gatacttcac ttcaggacgg
     9061 ttgtctgatc tacaacgtca agatcagggg ggtcaatttc acctcgaatg gaccagtaat
     9121 gcaaaaaaag accctgggat gggaagcttt tactgaaact ctctatcccg ccgacggcgg
     9181 cctcgaaggc agaaacgata tggctttgaa gctggttggc gggtcccatc taattgccaa
     9241 tgccaaaacc acatatagga gcaaaaagcc tgcaaagaac ctgaagatgc cgggtgtgta
     9301 ctatgttgac tatagattag agcggatcaa agaggcaaat aacgaaacgt acgtggagca
     9361 gcacgaggta gccgtggcgc gctactgcga tttaccctcc aaactgggcc ataaactgaa
     9421 ctgtacagga agcggagcta ctaacttcag cctgctgaag caggctggcg acgtggaaga
     9481 aaacccagga cctatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     9541 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     9601 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     9661 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     9721 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     9781 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     9841 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattcga
     9901 tatcaagctt atcggtaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     9961 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
    10021 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
    10081 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
    10141 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
    10201 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
    10261 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
    10321 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
    10381 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
    10441 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
    10501 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa
    10561 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
    10621 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
    10681 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
    10741 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
    10801 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
    10861 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
    10921 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
    10981 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
    11041 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    11101 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    11161 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
    11221 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    11281 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    11341 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    11401 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    11461 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
    11521 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    11581 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    11641 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    11701 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    11761 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    11821 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    11881 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    11941 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    12001 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    12061 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    12121 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    12181 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    12241 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    12301 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
    12361 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
    12421 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
    12481 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
    12541 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
    12601 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
    12661 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
    12721 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
    12781 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
    12841 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
    12901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    12961 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    13021 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
    13081 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    13141 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    13201 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    13261 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    13321 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    13381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    13441 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    13501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    13561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    13621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    13681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    13741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    13801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    13861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    13921 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    13981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    14041 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    14101 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    14161 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    14221 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    14281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    14341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    14401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    14461 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    14521 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    14581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    14641 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    14701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    14761 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    14821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    14881 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    14941 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    15001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    15061 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    15121 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    15181 caaatagggg ttccgcgcac atttccccga aaagtgcca
//
