LOCUS       sequence-457486-       14085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCAS9-VP64 with 2A Blast 
            resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14085)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 14085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: lenti dCAS-VP64_Blast
FEATURES             Location/Qualifiers
     source          1..14085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             26..46
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             71..220
                     /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             242..295
                     /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             296..694
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    719..1307
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(772..792)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1190..1201)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1310..1326)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1311..1327)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1832..2012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2038..2055)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2044..2268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2314..2742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2401..2420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2611..2632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2751..2771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2756..3085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2936..3071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2998..3017
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3133..3180
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3199..3573
                     /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    3703..3824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3740..3759)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3794..3813
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3873..3889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3873..3889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3886..3908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3897..3913
                     /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(3921..3927,3928..3945,3946..3951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3966..3987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4104..4121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4275..4863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4355..4374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5034..5825,5826..5894))
                     /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     5657..5676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5895..5999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6074..6093)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6265..6644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6646..6844
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6799..6819
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6862..7042
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7089..7214
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7707..7940
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8125..8169
                     /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    8467..8584
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8714..9892
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8945..9883
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     9840..9860
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             9908..9955
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(9956..9979,9980..9982,9983..12538,12539..12541,
                     12542..14056)
                     /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"
ORIGIN
        1 aggaagcgga ggaggaggta gcggacctaa gaaaaagagg aaggtggcgg ccgctggatc
       61 cggacgggct gacgcattgg acgattttga tctggatatg ctgggaagtg acgccctcga
      121 tgattttgac cttgacatgc ttggttcgga tgcccttgat gactttgacc tcgacatgct
      181 cggcagtgac gcccttgatg atttcgacct ggacatgctg attaactgta caggcagtgg
      241 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaatggc
      301 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
      361 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
      421 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
      481 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
      541 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
      601 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
      661 attgctgccc tctggttatg tgtgggaggg ctaagaattc gatatcaagc ttatcggtaa
      721 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
      781 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
      841 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
      901 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
      961 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     1021 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     1081 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     1141 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     1201 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     1261 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     1321 acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     1381 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     1441 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     1501 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     1561 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     1621 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     1681 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     1741 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     1801 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     1861 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     1921 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     1981 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
     2041 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2101 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2161 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2221 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2281 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2341 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2401 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2461 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2521 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2581 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2641 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2701 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2761 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2821 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2881 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2941 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3001 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3061 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3121 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     3181 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     3241 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     3301 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     3361 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     3421 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     3481 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3541 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     3601 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     3661 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     3721 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3781 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     3841 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3901 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3961 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4021 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5041 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5101 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5161 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5221 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5281 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5341 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5401 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5461 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5521 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5581 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5641 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5701 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5761 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5821 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5881 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5941 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6001 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     6061 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     6121 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     6181 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     6241 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     6301 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6361 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6421 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6481 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     6541 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     6601 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     6661 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     6721 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     6781 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     6841 gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6901 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6961 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7021 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     7081 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     7141 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     7201 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     7261 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7321 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7381 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7441 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7501 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7561 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     7621 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     7681 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     7741 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     7801 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     7861 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     7921 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     7981 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     8041 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     8101 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     8161 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     8221 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     8281 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     8341 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     8401 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat
     8461 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     8521 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     8581 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattagct agctgcaaag
     8641 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
     8701 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     8761 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     8821 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     8881 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     8941 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
     9001 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
     9061 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     9121 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     9181 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     9241 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     9301 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     9361 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     9421 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     9481 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     9541 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     9601 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     9661 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     9721 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     9781 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     9841 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gacgtacggc
     9901 caccatgaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggacaa
     9961 gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga
    10021 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
    10081 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
    10141 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
    10201 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
    10261 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
    10321 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
    10381 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
    10441 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
    10501 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
    10561 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
    10621 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc
    10681 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
    10741 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
    10801 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
    10861 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
    10921 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
    10981 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
    11041 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
    11101 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
    11161 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
    11221 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
    11281 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
    11341 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
    11401 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
    11461 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
    11521 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
    11581 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
    11641 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
    11701 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
    11761 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
    11821 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
    11881 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
    11941 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
    12001 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
    12061 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
    12121 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
    12181 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
    12241 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
    12301 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
    12361 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
    12421 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc acatcgtgcc
    12481 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaaggc
    12541 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
    12601 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
    12661 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
    12721 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
    12781 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
    12841 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
    12901 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
    12961 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
    13021 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
    13081 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
    13141 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
    13201 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
    13261 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
    13321 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
    13381 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
    13441 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
    13501 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
    13561 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
    13621 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
    13681 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
    13741 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
    13801 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
    13861 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
    13921 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
    13981 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
    14041 tcagctggga ggcgacagcg ctggaggagg tggaagcgga ggagg
//
