LOCUS       Exported               13632 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentivirus compatible dCas9-VPR construct driven by the EF1a 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    lenti EF1a-FLAG-dCas9-VPR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13632)
  AUTHORS   Savell KE, Bach SV, Zipperly ME, Revanna JS, Goska NA, Tuscher JJ, 
            Duke CG, Sultan FA, Burke JN, Williams D, Ianov L, Day JJ
  TITLE     A Neuron-Optimized CRISPR/dCas9 Activation System for Robust and 
            Specific Gene Regulation.
  JOURNAL   eNeuro. 2019 Mar 7;6(1). pii: eN-MNT-0495-18. doi: 
            10.1523/ENEURO.0495-18.2019. eCollection 2019 Jan-Feb.
  PUBMED    30863790
REFERENCE   2  (bases 1 to 13632)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13632
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    16..604
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(69..89)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(487..498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             676..909
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(935..952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    941..1165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1211..1639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1298..1317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1508..1529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1648..1668)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1653..1982
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1833..1968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1895..1914
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2030..2077
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2096..2470
                     /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    2600..2721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2637..2656)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2691..2710
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2770..2786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2770..2786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2783..2805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2794..2810
                     /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(2818..2848)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2863..2884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3001..3018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3172..3760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3252..3271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3931..4791)
                     /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     4554..4573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4792..4896)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4971..4990)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5162..5541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5543..5741
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5696..5716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5759..5939
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5986..6111
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6604..6837
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7022..7066
                     /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    7364..7481
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7532..7743
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7768..7833
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7840..7860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7885..11985
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11986..12033
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12061..12210
                     /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             12235..12255
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12685..13041
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             13060..13629
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
ORIGIN
        1 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
       61 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
      121 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
      181 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
      241 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
      301 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
      361 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
      421 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
      481 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
      541 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
      601 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
      661 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
      721 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
      781 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
      841 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
      901 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
      961 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1021 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1081 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1141 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     1201 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1261 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1321 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1381 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1441 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1501 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1561 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1621 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     1681 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     1741 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1801 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     1861 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     1921 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     1981 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     2041 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     2101 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     2161 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     2221 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     2281 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     2341 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     2401 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     2461 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     2521 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     2581 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2641 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2701 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     2761 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2821 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2881 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2941 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4921 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     4981 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5041 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     5101 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     5161 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     5221 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     5281 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     5341 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     5401 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     5461 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     5521 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     5581 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     5641 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     5701 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
     5761 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     5821 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     5881 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     5941 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     6001 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     6061 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     6121 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     6181 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     6241 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     6301 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     6361 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     6421 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     6481 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     6541 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     6601 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     6661 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     6721 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     6781 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     6841 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     6901 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     6961 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     7021 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     7081 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     7141 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     7201 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     7261 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     7321 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     7381 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     7441 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     7501 gcagagatcc agtttggtta attaaggtac cgggcagagc gcacatcgcc cacagtcccc
     7561 gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta
     7621 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     7681 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     7741 caggaccggt tctagagcgc tgccaccgac tataaggacc acgacggaga ctacaaggat
     7801 catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
     7861 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctggc catcggcacc
     7921 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     7981 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     8041 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     8101 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     8161 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     8221 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     8281 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     8341 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     8401 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     8461 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     8521 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     8581 aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     8641 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     8701 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     8761 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     8821 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     8881 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     8941 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     9001 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     9061 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     9121 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     9181 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     9241 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     9301 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     9361 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     9421 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     9481 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     9541 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     9601 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     9661 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     9721 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     9781 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     9841 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     9901 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     9961 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
    10021 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
    10081 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
    10141 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
    10201 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
    10261 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
    10321 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
    10381 tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgac
    10441 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
    10501 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
    10561 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
    10621 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
    10681 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
    10741 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
    10801 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
    10861 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
    10921 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
    10981 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
    11041 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
    11101 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
    11161 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
    11221 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
    11281 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
    11341 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
    11401 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
    11461 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
    11521 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
    11581 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
    11641 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
    11701 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
    11761 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
    11821 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
    11881 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
    11941 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
    12001 acgaaaaagg ccggacaggc caaaaagaaa aagctcgagg gcggaggcgg gagcggatcc
    12061 gacgcattgg acgattttga tctggatatg ctgggaagtg acgccctcga tgattttgac
    12121 cttgacatgc ttggttcgga tgcccttgat gactttgacc tcgacatgct cggcagtgac
    12181 gcccttgatg atttcgacct ggacatgctg attaactcta gaagttccgg atctccgaaa
    12241 aagaaacgca aagttggtag ccagtacctg cccgacaccg acgaccggca ccggatcgag
    12301 gaaaagcgga agcggaccta cgagacattc aagagcatca tgaagaagtc ccccttcagc
    12361 ggccccaccg accctagacc tccacctaga agaatcgccg tgcccagcag atccagcgcc
    12421 agcgtgccaa aacctgcccc ccagccttac cccttcacca gcagcctgag caccatcaac
    12481 tacgacgagt tccctaccat ggtgttcccc agcggccaga tctctcaggc ctctgctctg
    12541 gctccagccc ctcctcaggt gctgcctcag gctcctgctc ctgcaccagc tccagccatg
    12601 gtgtctgcac tggctcaggc accagcaccc gtgcctgtgc tggctcctgg acctccacag
    12661 gctgtggctc caccagcccc taaacctaca caggccggcg agggcacact gtctgaagct
    12721 ctgctgcagc tgcagttcga cgacgaggat ctgggagccc tgctgggaaa cagcaccgat
    12781 cctgccgtgt tcaccgacct ggccagcgtg gacaacagcg agttccagca gctgctgaac
    12841 cagggcatcc ctgtggcccc tcacaccacc gagcccatgc tgatggaata ccccgaggcc
    12901 atcacccggc tcgtgacagg cgctcagagg cctcctgatc cagctcctgc ccctctggga
    12961 gcaccaggcc tgcctaatgg actgctgtct ggcgacgagg acttcagctc tatcgccgat
    13021 atggatttct cagccttgct gggctctggc agcggcagcc gggattccag ggaagggatg
    13081 tttttgccga agcctgaggc cggctccgct attagtgacg tgtttgaggg ccgcgaggtg
    13141 tgccagccaa aacgaatccg gccatttcat cctccaggaa gtccatgggc caaccgccca
    13201 ctccccgcca gcctcgcacc aacaccaacc ggtccagtac atgagccagt cgggtcactg
    13261 accccggcac cagtccctca gccactggat ccagcgcccg cagtgactcc cgaggccagt
    13321 cacctgttgg aggatcccga tgaagagacg agccaggctg tcaaagccct tcgggagatg
    13381 gccgatactg tgattcccca gaaggaagag gctgcaatct gtggccaaat ggacctttcc
    13441 catccgcccc caaggggcca tctggatgag ctgacaacca cacttgagtc catgaccgag
    13501 gatctgaacc tggactcacc cctgaccccg gaattgaacg agattctgga taccttcctg
    13561 aacgacgagt gcctcttgca tgccatgcat atcagcacag gactgtccat cttcgacaca
    13621 tctctgtttt ga
//
