LOCUS       sequence-191019-       14292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCas9-VP64 with 2A puromycin 
            resistance marker (EF1a-dCas9-VP64-T2A-Puro-WPRE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14292)
  AUTHORS   Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I, 
            Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ
  TITLE     Evaluating Synthetic Activation and Repression of 
            Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and 
            Astrocytes.
  JOURNAL   Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 
            10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27.
  PUBMED    28757163
REFERENCE   2  (bases 1 to 14292)
  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"; doi: 
            "10.1016/j.stemcr.2017.06.012"; journalName: "Stem Cell Reports"; 
            date: "2017-08-8- 8"; volume: "9"; issue: "2"; pages: "615-628"
COMMENT     Sequence Label: lenti-EF1a-dCas9-VP64-Puro
FEATURES             Location/Qualifiers
     source          1..14292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    201..434
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             619..663
                     /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    961..1078
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1208..2386
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1439..2377
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2334..2354
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2402..2449
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(2450..2473,2474..2476,2477..5032,5033..5035,
                     5036..6550)
                     /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             6605..6625
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6650..6799
                     /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             6821..6874
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6887..7480
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="EYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVT
                     ELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQ
                     QQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAP
                     RNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7377..7397
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7505..8093
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7558..7578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7976..7987)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8096..8112)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8097..8113)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8618..8798
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8824..8841)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8830..9054
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9100..9528
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9187..9206)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9397..9418
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9537..9557)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9542..9871
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9722..9857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9784..9803
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9919..9966
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9985..10359
                     /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    10489..10610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10526..10545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10580..10599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10659..10675)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10659..10675)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10672..10694)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10683..10699
                     /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(10707..10713,10714..10731,10732..10737))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10752..10773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10890..10907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11061..11649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11141..11160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11820..12611,12612..12680))
                     /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     12443..12462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12681..12785)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12860..12879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13051..13430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        13432..13630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     13585..13605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             13648..13828
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    13875..14000
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
       61 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
      121 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
      181 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
      241 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
      301 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
      361 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
      421 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
      481 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
      541 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
      601 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
      661 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
      721 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
      781 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
      841 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
      901 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
      961 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     1021 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     1081 ggtttattac agggacagca gagatccagt ttggttaatt agctagctgc aaagatggat
     1141 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt
     1201 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     1261 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
     1321 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     1381 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
     1441 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
     1501 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     1561 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     1621 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     1681 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     1741 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     1801 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     1861 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     1921 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     1981 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     2041 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     2101 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     2161 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     2221 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     2281 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     2341 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat
     2401 gaaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg acaagaagta
     2461 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     2521 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     2581 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     2641 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     2701 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     2761 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     2821 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     2881 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     2941 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     3001 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     3061 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     3121 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
     3181 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     3241 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     3301 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     3361 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     3421 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     3481 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     3541 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     3601 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     3661 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     3721 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     3781 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     3841 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     3901 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     3961 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     4021 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     4081 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     4141 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     4201 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     4261 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     4321 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     4381 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     4441 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     4501 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     4561 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     4621 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     4681 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     4741 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     4801 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     4861 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     4921 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccacatcg tgcctcagag
     4981 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca aggcccgggg
     5041 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     5101 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     5161 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     5221 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     5281 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     5341 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     5401 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     5461 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     5521 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     5581 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     5641 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     5701 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     5761 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     5821 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     5881 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     5941 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     6001 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     6061 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     6121 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     6181 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     6241 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     6301 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     6361 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     6421 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     6481 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     6541 gggaggcgac agcgctggag gaggtggaag cggaggagga ggaagcggag gaggaggtag
     6601 cggacctaag aaaaagagga aggtggcggc cgctggatcc ggacgggctg acgcattgga
     6661 cgattttgat ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct
     6721 tggttcggat gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga
     6781 tttcgacctg gacatgctga ttaactgtac aggttcaggg gagggaagag gttccctgct
     6841 cacatgcgga gacgtagaag agaatcccgg acccatgacc tctagagagt acaagcccac
     6901 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     6961 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     7021 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     7081 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7141 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7201 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7261 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7321 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7381 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7441 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gaattcgata tcaagcttat
     7501 cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     7561 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     7621 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     7681 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     7741 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     7801 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     7861 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     7921 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     7981 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     8041 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     8101 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     8161 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     8221 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     8281 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     8341 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
     8401 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
     8461 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
     8521 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
     8581 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
     8641 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     8701 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     8761 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     8821 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     8881 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     8941 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     9001 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     9061 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     9121 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     9181 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     9241 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     9301 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     9361 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     9421 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     9481 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     9541 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     9601 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     9661 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     9721 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     9781 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     9841 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     9901 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     9961 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
    10021 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
    10081 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
    10141 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
    10201 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
    10261 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
    10321 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
    10381 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
    10441 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
    10501 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
    10561 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
    10621 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
    10681 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
    10741 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
    10801 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
    10861 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
    10921 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
    10981 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
    11041 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
    11101 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
    11161 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
    11221 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
    11281 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
    11341 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
    11401 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
    11461 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
    11521 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
    11581 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    11641 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    11701 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    11761 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    11821 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    11881 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    11941 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    12001 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    12061 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    12121 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    12181 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    12241 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    12301 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    12361 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    12421 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    12481 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    12541 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    12601 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    12661 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    12721 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    12781 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
    12841 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
    12901 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
    12961 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
    13021 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
    13081 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
    13141 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
    13201 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
    13261 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
    13321 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
    13381 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
    13441 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
    13501 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
    13561 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
    13621 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
    13681 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    13741 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    13801 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
    13861 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
    13921 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
    13981 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
    14041 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
    14101 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
    14161 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
    14221 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
    14281 aagctttaga ca
//
