LOCUS       sequence-221563-       14227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus compatible dCas9-VPR construct driven by the CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14227)
  AUTHORS   Savell KE, Bach SV, Zipperly ME, Revanna JS, Goska NA, Tuscher JJ, 
            Duke CG, Sultan FA, Burke JN, Williams D, Ianov L, Day JJ
  TITLE     A Neuron-Optimized CRISPR/dCas9 Activation System for Robust and 
            Specific Gene Regulation.
  JOURNAL   eNeuro. 2019 Mar 7;6(1). pii: eN-MNT-0495-18. doi: 
            10.1523/ENEURO.0495-18.2019. eCollection 2019 Jan-Feb.
  PUBMED    30863790
REFERENCE   2  (bases 1 to 14227)
  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: "eNeuro."; 
            date: "2019-03-7"; volume: "6(1). pii: eN-MNT-0495-18. doi"; pages: 
            " 10.1523/ENEURO.0495-18.2019. eCollection 2019 Jan-Feb"
COMMENT     Sequence Label: lenti CAG-FLAG-dCas9-VPR
FEATURES             Location/Qualifiers
     source          1..14227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(166..957,958..1026))
                     /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     789..808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1027..1131)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1206..1225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1397..1776
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1778..1976
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1931..1951
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1994..2174
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2221..2346
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2839..3072
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3257..3301
                     /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    3599..3716
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3767..4052
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          4332..4559
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      4571..4580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4580..4645
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4652..4672
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4697..4720,4721..4723,4724..7210,7211..7213,
                     7214..8797)
                     /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 H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4697..8797
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8798..8845
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8873..9022
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             9047..9067
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9497..9853
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             9872..10441
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     misc_feature    10478..11066
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10531..10551)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10949..10960)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11138..11371
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11397..11414)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11403..11627
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11673..12101
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11760..11779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11970..11991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12110..12130)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12115..12444
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12295..12430
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12357..12376
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12492..12539
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12558..12932
                     /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    13062..13183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13099..13118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13153..13172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13232..13248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13232..13248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13245..13267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13256..13272
                     /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(13280..13286,13287..13304,13305..13310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13325..13346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13463..13480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13634..14222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13714..13733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
       61 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
      301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
      361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
      421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
      481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
      541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
      601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
      661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
      721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
      781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
      841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
      901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
      961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1141 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     1201 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     1261 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     1321 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     1381 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     1441 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     1501 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1561 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1621 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1681 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1741 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1801 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1861 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1921 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg
     1981 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     2041 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     2101 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     2161 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     2221 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     2281 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     2341 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     2401 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     2461 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     2521 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     2581 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     2641 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     2701 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     2761 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     2821 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     2881 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     2941 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     3001 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     3061 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     3121 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     3181 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     3241 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     3301 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     3361 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     3421 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     3481 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     3541 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
     3601 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     3661 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     3721 tttattacag ggacagcaga gatccagttt ggttaattaa ggtacccgtt acataactta
     3781 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
     3841 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     3901 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     3961 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
     4021 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     4081 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     4141 tttgtgcagc gatgggggcg gggggggggg gggggsgsgs gccrgggggg gggggggggg
     4201 ggggggsgsg gggsggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     4261 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     4321 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     4381 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     4441 gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg
     4501 ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg
     4561 ttggaccggt gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat
     4621 tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca
     4681 cggagtccca gcagccgaca agaagtacag catcggcctg gccatcggca ccaactctgt
     4741 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     4801 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     4861 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     4921 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     4981 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     5041 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     5101 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     5161 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     5221 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     5281 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     5341 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     5401 cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     5461 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     5521 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     5581 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     5641 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     5701 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     5761 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     5821 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     5881 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     5941 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     6001 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     6061 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     6121 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     6181 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     6241 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     6301 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     6361 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     6421 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     6481 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     6541 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     6601 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     6661 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     6721 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     6781 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     6841 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     6901 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     6961 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     7021 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     7081 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     7141 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     7201 cgatgtggac gctatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     7261 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     7321 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     7381 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     7441 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     7501 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     7561 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     7621 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     7681 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     7741 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     7801 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     7861 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     7921 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     7981 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     8041 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     8101 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     8161 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     8221 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     8281 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     8341 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     8401 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     8461 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     8521 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     8581 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     8641 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     8701 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     8761 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
     8821 ggccggacag gccaaaaaga aaaagctcga gggcggaggc gggagcggat ccgacgcatt
     8881 ggacgatttt gatctggata tgctgggaag tgacgccctc gatgattttg accttgacat
     8941 gcttggttcg gatgcccttg atgactttga cctcgacatg ctcggcagtg acgcccttga
     9001 tgatttcgac ctggacatgc tgattaactc tagaagttcc ggatctccga aaaagaaacg
     9061 caaagttggt agccagtacc tgcccgacac cgacgaccgg caccggatcg aggaaaagcg
     9121 gaagcggacc tacgagacat tcaagagcat catgaagaag tcccccttca gcggccccac
     9181 cgaccctaga cctccaccta gaagaatcgc cgtgcccagc agatccagcg ccagcgtgcc
     9241 aaaacctgcc ccccagcctt accccttcac cagcagcctg agcaccatca actacgacga
     9301 gttccctacc atggtgttcc ccagcggcca gatctctcag gcctctgctc tggctccagc
     9361 ccctcctcag gtgctgcctc aggctcctgc tcctgcacca gctccagcca tggtgtctgc
     9421 actggctcag gcaccagcac ccgtgcctgt gctggctcct ggacctccac aggctgtggc
     9481 tccaccagcc cctaaaccta cacaggccgg cgagggcaca ctgtctgaag ctctgctgca
     9541 gctgcagttc gacgacgagg atctgggagc cctgctggga aacagcaccg atcctgccgt
     9601 gttcaccgac ctggccagcg tggacaacag cgagttccag cagctgctga accagggcat
     9661 ccctgtggcc cctcacacca ccgagcccat gctgatggaa taccccgagg ccatcacccg
     9721 gctcgtgaca ggcgctcaga ggcctcctga tccagctcct gcccctctgg gagcaccagg
     9781 cctgcctaat ggactgctgt ctggcgacga ggacttcagc tctatcgccg atatggattt
     9841 ctcagccttg ctgggctctg gcagcggcag ccgggattcc agggaaggga tgtttttgcc
     9901 gaagcctgag gccggctccg ctattagtga cgtgtttgag ggccgcgagg tgtgccagcc
     9961 aaaacgaatc cggccatttc atcctccagg aagtccatgg gccaaccgcc cactccccgc
    10021 cagcctcgca ccaacaccaa ccggtccagt acatgagcca gtcgggtcac tgaccccggc
    10081 accagtccct cagccactgg atccagcgcc cgcagtgact cccgaggcca gtcacctgtt
    10141 ggaggatccc gatgaagaga cgagccaggc tgtcaaagcc cttcgggaga tggccgatac
    10201 tgtgattccc cagaaggaag aggctgcaat ctgtggccaa atggaccttt cccatccgcc
    10261 cccaaggggc catctggatg agctgacaac cacacttgag tccatgaccg aggatctgaa
    10321 cctggactca cccctgaccc cggaattgaa cgagattctg gataccttcc tgaacgacga
    10381 gtgcctcttg catgccatgc atatcagcac aggactgtcc atcttcgaca catctctgtt
    10441 ttgagaattc aattctaact agacgcgtta agtcgacaat caacctctgg attacaaaat
    10501 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
    10561 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
    10621 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
    10681 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
    10741 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
    10801 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
    10861 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
    10921 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
    10981 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
    11041 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
    11101 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
    11161 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
    11221 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
    11281 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    11341 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
    11401 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
    11461 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
    11521 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
    11581 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
    11641 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
    11701 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
    11761 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
    11821 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
    11881 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
    11941 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
    12001 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
    12061 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
    12121 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    12181 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
    12241 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
    12301 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
    12361 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
    12421 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
    12481 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
    12541 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
    12601 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
    12661 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
    12721 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
    12781 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
    12841 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
    12901 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
    12961 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
    13021 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
    13081 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    13141 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    13201 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    13261 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    13321 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    13381 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    13441 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    13501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    13561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    13621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    13681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    13741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    13801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    13861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    13921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    13981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    14041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    14101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    14161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    14221 aagaaga
//
