LOCUS       sequence-215531-       14228 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 14228)
  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 14228)
  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..14228
                     /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"
     promoter        4054..4331
                     /label=chicken beta-actin promoter
     intron          4332..4560
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      4572..4581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4581..4646
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4653..4673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4698..4721,4722..4724,4725..7211,7212..7214,
                     7215..8798)
                     /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             4698..8798
                     /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             8799..8846
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8874..9023
                     /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             9048..9068
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9498..9854
                     /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             9873..10442
                     /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    10479..11067
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10532..10552)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10950..10961)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11139..11372
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11398..11415)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11404..11628
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11674..12102
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11761..11780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11971..11992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12111..12131)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12116..12445
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12296..12431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12358..12377
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12493..12540
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12559..12933
                     /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    13063..13184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13100..13119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13154..13173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13233..13249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13233..13249)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13246..13268)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13257..13273
                     /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(13281..13287,13288..13305,13306..13311))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13326..13347
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13464..13481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13635..14223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13715..13734)
                     /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 gggggcgcgc gccaggcggg gcggggcggg
     4201 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     4261 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     4321 cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc cgccgccgcc
     4381 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
     4441 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
     4501 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
     4561 gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag gatcatgata
     4621 ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc
     4681 acggagtccc agcagccgac aagaagtaca gcatcggcct ggccatcggc accaactctg
     4741 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     4801 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     4861 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     4921 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     4981 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     5041 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     5101 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     5161 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     5221 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     5281 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     5341 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     5401 gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     5461 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     5521 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     5581 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     5641 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     5701 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     5761 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     5821 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     5881 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     5941 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     6001 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     6061 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     6121 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     6181 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     6241 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     6301 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     6361 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     6421 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     6481 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     6541 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     6601 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     6661 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     6721 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     6781 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     6841 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     6901 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     6961 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     7021 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     7081 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     7141 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     7201 acgatgtgga cgctatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     7261 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     7321 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     7381 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     7441 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     7501 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     7561 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     7621 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     7681 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     7741 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     7801 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     7861 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     7921 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     7981 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     8041 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     8101 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     8161 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     8221 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     8281 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     8341 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     8401 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     8461 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     8521 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     8581 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     8641 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     8701 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     8761 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
     8821 aggccggaca ggccaaaaag aaaaagctcg agggcggagg cgggagcgga tccgacgcat
     8881 tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca
     8941 tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg
     9001 atgatttcga cctggacatg ctgattaact ctagaagttc cggatctccg aaaaagaaac
     9061 gcaaagttgg tagccagtac ctgcccgaca ccgacgaccg gcaccggatc gaggaaaagc
     9121 ggaagcggac ctacgagaca ttcaagagca tcatgaagaa gtcccccttc agcggcccca
     9181 ccgaccctag acctccacct agaagaatcg ccgtgcccag cagatccagc gccagcgtgc
     9241 caaaacctgc cccccagcct taccccttca ccagcagcct gagcaccatc aactacgacg
     9301 agttccctac catggtgttc cccagcggcc agatctctca ggcctctgct ctggctccag
     9361 cccctcctca ggtgctgcct caggctcctg ctcctgcacc agctccagcc atggtgtctg
     9421 cactggctca ggcaccagca cccgtgcctg tgctggctcc tggacctcca caggctgtgg
     9481 ctccaccagc ccctaaacct acacaggccg gcgagggcac actgtctgaa gctctgctgc
     9541 agctgcagtt cgacgacgag gatctgggag ccctgctggg aaacagcacc gatcctgccg
     9601 tgttcaccga cctggccagc gtggacaaca gcgagttcca gcagctgctg aaccagggca
     9661 tccctgtggc ccctcacacc accgagccca tgctgatgga ataccccgag gccatcaccc
     9721 ggctcgtgac aggcgctcag aggcctcctg atccagctcc tgcccctctg ggagcaccag
     9781 gcctgcctaa tggactgctg tctggcgacg aggacttcag ctctatcgcc gatatggatt
     9841 tctcagcctt gctgggctct ggcagcggca gccgggattc cagggaaggg atgtttttgc
     9901 cgaagcctga ggccggctcc gctattagtg acgtgtttga gggccgcgag gtgtgccagc
     9961 caaaacgaat ccggccattt catcctccag gaagtccatg ggccaaccgc ccactccccg
    10021 ccagcctcgc accaacacca accggtccag tacatgagcc agtcgggtca ctgaccccgg
    10081 caccagtccc tcagccactg gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt
    10141 tggaggatcc cgatgaagag acgagccagg ctgtcaaagc ccttcgggag atggccgata
    10201 ctgtgattcc ccagaaggaa gaggctgcaa tctgtggcca aatggacctt tcccatccgc
    10261 ccccaagggg ccatctggat gagctgacaa ccacacttga gtccatgacc gaggatctga
    10321 acctggactc acccctgacc ccggaattga acgagattct ggataccttc ctgaacgacg
    10381 agtgcctctt gcatgccatg catatcagca caggactgtc catcttcgac acatctctgt
    10441 tttgagaatt caattctaac tagacgcgtt aagtcgacaa tcaacctctg gattacaaaa
    10501 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
    10561 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
    10621 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
    10681 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
    10741 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
    10801 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
    10861 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
    10921 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
    10981 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
    11041 ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc
    11101 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    11161 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
    11221 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    11281 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    11341 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
    11401 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
    11461 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
    11521 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
    11581 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
    11641 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
    11701 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
    11761 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
    11821 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
    11881 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
    11941 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
    12001 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
    12061 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
    12121 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
    12181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
    12241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
    12301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
    12361 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
    12421 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
    12481 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
    12541 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
    12601 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
    12661 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
    12721 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
    12781 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
    12841 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
    12901 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
    12961 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
    13021 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
    13081 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
    13141 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
    13201 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    13261 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    13321 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    13381 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    13441 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    13501 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    13561 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    13621 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    13681 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    13741 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    13801 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    13861 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    13921 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    13981 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    14041 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    14101 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    14161 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    14221 caagaaga
//
