LOCUS       sequence-369806-       14805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 expression vector with cloning site for C terminal fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14805)
  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 14805)
  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: pJT216 Lenti
            pEF-dCas9-3XFLAG-LibCloneSite-T2A...
FEATURES             Location/Qualifiers
     source          1..14805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        221..600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        602..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     755..775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             818..998
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1045..1170
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1663..1896
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2081..2125
                     /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    2423..2540
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2670..3848
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2901..3839
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3796..3816
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3864..3911
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3912..3935,3936..3938,3939..6494,6495..6497,
                     6498..8012)
                     /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             8067..8087
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8103..8168
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8220..8273
                     /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             9000..9056
                     /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             9057..9455
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9480..10068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9533..9553)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9951..9962)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10071..10087)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10072..10088)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10593..10773
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10799..10816)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10805..11029
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11075..11503
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11162..11181)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11372..11393
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11512..11532)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11517..11846
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11697..11832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11759..11778
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11894..11941
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11960..12334
                     /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    12464..12585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12501..12520)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12555..12574
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12634..12650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12634..12650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12647..12669)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12658..12674
                     /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(12682..12688,12689..12706,12707..12712))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12727..12748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12865..12882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13036..13624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13116..13135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13795..14586,14587..14655))
                     /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     14418..14437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14656..14760)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
       61 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      121 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      181 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      241 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      301 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      361 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      421 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      481 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      541 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      601 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      661 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      721 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      781 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
      841 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      901 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
      961 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     1021 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     1081 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     1141 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     1201 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     1261 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     1321 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     1381 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     1441 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     1501 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1561 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1621 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1681 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1741 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1801 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1861 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1921 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1981 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2041 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2101 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2161 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2221 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2281 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     2341 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2401 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2461 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2521 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
     2581 ttagctagct gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga
     2641 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
     2701 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     2761 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2821 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2881 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     2941 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
     3001 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
     3061 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
     3121 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
     3181 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
     3241 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
     3301 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
     3361 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
     3421 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
     3481 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
     3541 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
     3601 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     3661 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     3721 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     3781 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     3841 tgtcgtgacg tacggccacc atgaaaaggc cggcggccac gaaaaaggcc ggccaggcaa
     3901 aaaagaaaaa ggacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct
     3961 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     4021 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     4081 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     4141 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     4201 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     4261 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     4321 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     4381 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     4441 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     4501 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     4561 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     4621 tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     4681 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     4741 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     4801 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     4861 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     4921 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     4981 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     5041 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     5101 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     5161 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     5221 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     5281 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     5341 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     5401 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     5461 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     5521 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     5581 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     5641 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     5701 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     5761 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     5821 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     5881 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     5941 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     6001 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     6061 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     6121 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     6181 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     6241 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     6301 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     6361 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     6421 tggaccacat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     6481 ccagaagcga caaggcccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     6541 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     6601 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     6661 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     6721 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     6781 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     6841 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     6901 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     6961 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     7021 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     7081 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     7141 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     7201 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     7261 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     7321 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     7381 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     7441 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     7501 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     7561 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     7621 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     7681 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     7741 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     7801 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     7861 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     7921 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     7981 cacggatcga cctgtctcag ctgggaggcg acagcgctgg aggaggtgga agcggaggag
     8041 gaggaagcgg aggaggaggt agcggaccta agaaaaagag gaaggtggcg gccgctggat
     8101 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     8161 atgacaaggg tggatccgga tcaggctccg gagacgggtc aagcgtctct ggatctggag
     8221 agggaagggg aagcctccta acttgcggag atgtcgagga gaatcctggc ccaatggtta
     8281 gtaagggaga ggagttgata aaagagaata tgcacatgaa actgtatatg gaagggacag
     8341 tggacaacca tcacttcaag tgcacctctg agggagaagg aaagccatac gaaggcaccc
     8401 agactatgcg aattaaggtc gtggagggag ggcctctccc cttcgccttc gatattcttg
     8461 caacatcttt cctctacggg agcaaaacct tcatcaatca cacacaaggc atacctgact
     8521 tttttaagca gagttttcca gagggcttta cctgggagcg tgtgactaca tacgaagatg
     8581 ggggtgtgct gacggctaca caggatactt cacttcagga cggttgtctg atctacaacg
     8641 tcaagatcag gggggtcaat ttcacctcga atggaccagt aatgcaaaaa aagaccctgg
     8701 gatgggaagc ttttactgaa actctctatc ccgccgacgg cggcctcgaa ggcagaaacg
     8761 atatggcttt gaagctggtt ggcgggtccc atctaattgc caatgccaaa accacatata
     8821 ggagcaaaaa gcctgcaaag aacctgaaga tgccgggtgt gtactatgtt gactatagat
     8881 tagagcggat caaagaggca aataacgaaa cgtacgtgga gcagcacgag gtagccgtgg
     8941 cgcgctactg cgatttaccc tccaaactgg gccataaact gaactgtaca ggaagcggag
     9001 ctactaactt cagcctgctg aagcaggctg gcgacgtgga agaaaaccca ggacctatgg
     9061 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     9121 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     9181 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     9241 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     9301 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     9361 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     9421 aattgctgcc ctctggttat gtgtgggagg gctaagaatt cgatatcaag cttatcggta
     9481 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     9541 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     9601 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     9661 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     9721 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     9781 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     9841 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     9901 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     9961 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
    10021 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
    10081 gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
    10141 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
    10201 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
    10261 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
    10321 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
    10381 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
    10441 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
    10501 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
    10561 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
    10621 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
    10681 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    10741 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
    10801 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
    10861 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
    10921 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
    10981 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
    11041 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
    11101 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
    11161 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
    11221 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
    11281 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
    11341 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
    11401 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    11461 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
    11521 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    11581 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
    11641 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    11701 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
    11761 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
    11821 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
    11881 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
    11941 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
    12001 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
    12061 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
    12121 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
    12181 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
    12241 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
    12301 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
    12361 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
    12421 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
    12481 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    12541 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
    12601 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    12661 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    12721 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    12781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    12841 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    12901 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    12961 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    13021 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    13081 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    13141 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    13201 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    13261 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    13321 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    13381 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    13441 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    13501 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    13561 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    13621 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    13681 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    13741 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    13801 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    13861 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    13921 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
    13981 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    14041 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    14101 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    14161 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    14221 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    14281 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    14341 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    14401 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    14461 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    14521 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    14581 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    14641 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    14701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    14761 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggaga
//
