pLentiV2-dCas9-VP64

LOCUS       pLentiV2-dCas9-VP64              15520 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of gRNA and dCas9-VP64 from the same plasmid for CRISPRa.
ACCESSION   .
VERSION     .
KEYWORDS    pLentiV2-dCas9-VP64
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15520)
  TITLE     pLentiV2-dCas9-VP64
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 15520)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..15520
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2611
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4501..4576
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4638..4849
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4866..4913
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4914..9014
                     /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             9069..9089
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9114..9263
                     /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             9285..9341
                     /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             9342..9938
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9835..9855
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9975..10563
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10028..10048)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10446..10457)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10566..10582)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10567..10583)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11088..11268
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11294..11311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11300..11524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11570..11998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11657..11676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11867..11888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12007..12027)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12012..12341
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12192..12327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12254..12273
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12389..12436
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12455..12829
                     /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    12959..13080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12996..13015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13050..13069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13129..13145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13129..13145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13142..13164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13153..13169
                     /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(13177..13207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13222..13243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13360..13377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13531..14119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13611..13630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14290..15150)
                     /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     14913..14932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15151..15255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(15330..15349)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagacggttg taaatgagca
     2641 cacaaaatac acatgctaaa atattatatt ctatgacctt tataaaatca accaaaatct
     2701 tctttttaat aactttagta tcaataatta gaatttttat gttccttttt gcaaactttt
     2761 aataaaaatg agcaaaataa aaaaacgcta gttttagtaa ctcgcgttgt tttcttcacc
     2821 tttaataata gctactccac cacttgttcc taagcggtca gctcctgctt caatcatttt
     2881 ttgagcatct tcaaatgttc taactccacc agctgcttta actaaagcat tgtctttaac
     2941 aactgacttc attagtttaa catcttcaaa tgttgcacct gattttgaaa atcctgttga
     3001 tgttttaaca aattctaatc cagcttcaac agctatttca caagctttca tgatttcttc
     3061 ttttgttaat aaacaatttt ccataataca tttaacaaca tgtgatccag ctgctttttt
     3121 tacagctttc atgtcttcta aaactaattc ataatttttg tcttttaatg caccaatatt
     3181 taataccata tcaatttctg ttgcaccatc tttaattgct tcagaaactt cgaatgcttt
     3241 tgtagctgtt gtgcatgcac ctagaggaaa acctacaaca tttgttattc ctacatttgt
     3301 gccttttaat aattctttac aatagcttgt tcaatatgaa ttaacacaaa ctgttgcaaa
     3361 atcaaattca attgcttcat cacataattg tttaatttca gctttcgtag catcttgttt
     3421 taataatgtg tgatctatat atttgtttag tttcattttt tctcctatat attcattttt
     3481 aattttaatt ctttaataat ttcgtctact ttaactttag cgttttgaac agattcacca
     3541 acacctataa aataaatttt tagtttaggt tcagttccac ttgggcgaac agcaaatcat
     3601 gacttatctt ctaaataaaa ttttagtaag tcttgtcctg gcatattata cattccatcg
     3661 atgtagtctt caacattaac aactttaagt ccagcaattt gagttaaggg tgttgctctc
     3721 aatgatttca ttaatggttc aatttttaat ttcttttctt ctggtttaaa attcaagttt
     3781 aaagtgaaag tgtaatatgc acccatttct ttaaataaat cttctaaata gtctactaat
     3841 gttttatttt gttttttata aaatcaagca gcctctgcta ttaatataga agcttgtatt
     3901 ccatctttat ctctagctga gtcatcaatt acatatccat aactttcttc ataagcaaaa
     3961 acaaaattta atccgttatc ttcttcttta gcaatttctc tacccattca tttaaatcca
     4021 gttaaagttt ttacaatatt aactccatat ttttcatgag cgattctatc acccaaatca
     4081 cttgttacaa aacttgaata tagagccgga ttttttggaa tgctatttaa gcgttttaga
     4141 tttgataatt ttcaatcaat taaaattggt cctgtttgat ttccatctaa tcttacaaaa
     4201 tgaccatcat gttttattgc cattccaaat ctgtcagcat ctgggtcatt cataataata
     4261 atatctgcat catgtttaat accatattca agcggtattt ttcatgcagg atcaaattct
     4321 ggatttggat ttacaacatt tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca
     4381 ataacgttat atcctgattc acgtaatatt tttggggtaa atttagttcc tgttccatta
     4441 actgcgctaa aaataatttt taaatctttt ttagcttctt gctctttttt gtacgtctct
     4501 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     4561 ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg
     4621 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4681 aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4741 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4801 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg cgtacggcca
     4861 ccatgaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga
     4921 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg
     4981 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     5041 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     5101 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     5161 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     5221 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     5281 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     5341 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     5401 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     5461 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     5521 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     5581 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc
     5641 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     5701 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     5761 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     5821 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     5881 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     5941 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     6001 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     6061 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     6121 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     6181 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     6241 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     6301 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     6361 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     6421 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     6481 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     6541 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     6601 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     6661 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     6721 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     6781 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     6841 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     6901 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     6961 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     7021 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     7081 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     7141 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     7201 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     7261 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     7321 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     7381 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc
     7441 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc
     7501 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     7561 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     7621 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     7681 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     7741 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     7801 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     7861 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     7921 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     7981 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     8041 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     8101 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     8161 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     8221 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     8281 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     8341 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     8401 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     8461 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     8521 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     8581 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     8641 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     8701 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     8761 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     8821 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     8881 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     8941 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     9001 agctgggagg cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag
     9061 gtagcggacc taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat
     9121 tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca
     9181 tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg
     9241 atgatttcga cctggacatg ctgattaact gtacaggcag tggagcaaca aacttctctc
     9301 tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg
     9361 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     9421 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     9481 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     9541 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     9601 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     9661 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     9721 tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     9781 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     9841 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     9901 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gaattcgata
     9961 tcaagcttat cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
    10021 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
    10081 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
    10141 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
    10201 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
    10261 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
    10321 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
    10381 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
    10441 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
    10501 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
    10561 cgcatcgata ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata
    10621 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
    10681 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
    10741 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
    10801 atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca
    10861 caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag
    10921 ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg
    10981 tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc
    11041 gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt
    11101 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
    11161 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
    11221 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
    11281 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
    11341 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
    11401 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
    11461 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
    11521 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
    11581 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
    11641 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
    11701 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
    11761 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    11821 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
    11881 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
    11941 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
    12001 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
    12061 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
    12121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
    12181 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
    12241 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
    12301 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
    12361 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
    12421 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
    12481 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
    12541 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
    12601 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
    12661 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
    12721 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
    12781 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
    12841 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
    12901 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
    12961 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
    13021 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
    13081 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
    13141 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    13201 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    13261 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    13321 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    13381 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    13441 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    13501 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    13561 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    13621 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    13681 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    13741 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    13801 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    13861 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
    13921 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
    13981 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    14041 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    14101 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    14161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
    14221 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    14281 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    14341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    14401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    14461 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    14521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    14581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    14641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    14701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    14761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    14821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    14881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    14941 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    15001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    15061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    15121 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    15181 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    15241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
    15301 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
    15361 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
    15421 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
    15481 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//