LOCUS       sequence-252713-       14337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCas9-KRAB with 2A puromycin 
            resistance marker (EF1a-dCas9-KRAB-T2A-Puro-WPRE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14337)
  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 14337)
  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-KRAB-Puro
FEATURES             Location/Qualifiers
     source          1..14337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    179..412
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             597..641
                     /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    939..1056
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1186..2364
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1417..2355
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2312..2332
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2380..2427
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(2428..2451,2452..2454,2455..5010,5011..5013,
                     5014..6528)
                     /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             6583..6603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6628..6822
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             6844..6897
                     /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             6910..7503
                     /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     7400..7420
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7528..8116
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7581..7601)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7999..8010)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8119..8135)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8120..8136)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8641..8821
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8847..8864)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8853..9077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9123..9551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9210..9229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9420..9441
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9560..9580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9565..9894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9745..9880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9807..9826
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9942..9989
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10008..10382
                     /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    10512..10633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10549..10568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10603..10622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10682..10698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10682..10698)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10695..10717)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10706..10722
                     /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(10730..10736,10737..10754,10755..10760))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10775..10796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10913..10930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11084..11672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11164..11183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11843..12634,12635..12703))
                     /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     12466..12485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12704..12808)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12883..12902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13074..13453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        13455..13653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     13608..13628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             13671..13851
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    13898..14023
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
       61 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
      121 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
      181 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
      241 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
      301 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
      361 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
      421 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
      481 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
      541 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
      601 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
      661 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
      721 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
      781 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
      841 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
      901 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
      961 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     1021 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     1081 gatccagttt ggttaattag ctagctgcaa agatggataa agttttaaac agagaggaat
     1141 ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
     1201 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     1261 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     1321 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     1381 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
     1441 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
     1501 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
     1561 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     1621 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     1681 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     1741 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     1801 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     1861 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
     1921 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     1981 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     2041 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     2101 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     2161 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     2221 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     2281 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     2341 tttcttccat ttcaggtgtc gtgacgtacg gccaccatga aaaggccggc ggccacgaaa
     2401 aaggccggcc aggcaaaaaa gaaaaaggac aagaagtaca gcatcggcct ggccatcggc
     2461 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     2521 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     2581 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     2641 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     2701 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     2761 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     2821 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     2881 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     2941 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     3001 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     3061 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     3121 aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc
     3181 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     3241 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     3301 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     3361 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     3421 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     3481 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     3541 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     3601 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     3661 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     3721 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     3781 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     3841 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     3901 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     3961 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     4021 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     4081 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     4141 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     4201 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     4261 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     4321 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     4381 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     4441 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     4501 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     4561 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     4621 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     4681 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     4741 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     4801 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     4861 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     4921 ctgtccgact acgatgtgga ccacatcgtg cctcagagct ttctgaagga cgactccatc
     4981 gacaacaagg tgctgaccag aagcgacaag gcccggggca agagcgacaa cgtgccctcc
     5041 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     5101 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     5161 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     5221 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     5281 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     5341 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     5401 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     5461 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     5521 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     5581 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     5641 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     5701 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     5761 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     5821 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     5881 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     5941 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     6001 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     6061 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     6121 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     6181 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     6241 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     6301 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     6361 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     6421 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     6481 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacag cgctggagga
     6541 ggtggaagcg gaggaggagg aagcggagga ggaggtagcg gacctaagaa aaagaggaag
     6601 gtggcggccg ctggatccgg acgggctaga acactggtta cgttcaagga cgtgtttgtg
     6661 gactttacac gtgaggagtg gaaattgctg gatactgcgc aacaaattgt gtatcgaaat
     6721 gtcatgcttg agaattacaa gaacctcgtc agtctcggat accagttgac gaaaccggat
     6781 gtgatcctta ggctcgaaaa gggggaagaa ccttggctgg taattaactg tacaggttca
     6841 ggggagggaa gaggttccct gctcacatgc ggagacgtag aagagaatcc cggacccatg
     6901 acctctagag agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     6961 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7021 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7081 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     7141 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     7201 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     7261 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     7321 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     7381 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     7441 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     7501 tgagaattcg atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga
     7561 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     7621 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     7681 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     7741 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     7801 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     7861 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     7921 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     7981 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     8041 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     8101 tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc
     8161 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     8221 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     8281 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     8341 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     8401 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
     8461 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
     8521 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
     8581 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
     8641 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     8701 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     8761 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     8821 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     8881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     8941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     9001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     9061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     9121 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     9181 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     9241 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     9301 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     9361 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     9421 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     9481 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     9541 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     9601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     9661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     9721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     9781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     9841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     9901 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     9961 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
    10021 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
    10081 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
    10141 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
    10201 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
    10261 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
    10321 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
    10381 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
    10441 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
    10501 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    10561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    10621 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
    10681 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
    10741 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
    10801 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
    10861 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
    10921 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
    10981 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
    11041 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    11101 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    11161 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    11221 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    11281 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    11341 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    11401 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    11461 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
    11521 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
    11581 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
    11641 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
    11701 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
    11761 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
    11821 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
    11881 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
    11941 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
    12001 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
    12061 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
    12121 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
    12181 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
    12241 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    12301 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    12361 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    12421 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    12481 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    12541 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    12601 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    12661 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    12721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    12781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
    12841 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
    12901 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
    12961 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
    13021 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
    13081 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
    13141 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
    13201 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
    13261 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
    13321 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
    13381 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
    13441 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
    13501 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
    13561 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
    13621 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
    13681 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
    13741 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    13801 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
    13861 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
    13921 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
    13981 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
    14041 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
    14101 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
    14161 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
    14221 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
    14281 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaa
//
