LOCUS       Exported               14238 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses a SYN-driven, intron-containing dCas9-VPR fusion in which 
            the part of the dCas9 cassette is double floxed and in inverted 
            orientation (DIO). Requires Cre-dependent recombination..
ACCESSION   .
VERSION     .
KEYWORDS    lenti SYN-SVI-DIO-dCas9-VPR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14238)
  TITLE     A Cre-dependent CRISPR/dCas9 activation system for gene expression 
            regulation in neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14238)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14238
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(166..1026)
                     /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     789..808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1027..1131)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1206..1225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1397..1776
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1778..1976
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1931..1951
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1994..2174
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2221..2346
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2839..3072
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3257..3301
                     /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    3599..3716
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3761..4208
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    4221..4254
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    4305..4338
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(5826..5846)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5853..5918)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      5918..5927
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(5928..5961)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(6012..6045)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             6310..8808
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
                     PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             8809..8856
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8884..9033
                     /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             9058..9078
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9508..9864
                     /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             9883..10452
                     /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"
     misc_feature    10489..11077
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10542..10562)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10960..10971)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11149..11382
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11408..11425)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11414..11638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11684..12112
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11771..11790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11981..12002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12121..12141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12126..12455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12306..12441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12368..12387
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12503..12550
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12569..12943
                     /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    13073..13194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13110..13129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13164..13183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13243..13259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13243..13259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13256..13278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13267..13283
                     /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(13291..13321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13336..13357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13474..13491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13645..14233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13725..13744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
       61 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
      301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
      361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
      421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
      481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
      541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
      601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
      661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
      721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
      781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
      841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
      901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
      961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1141 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     1201 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     1261 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     1321 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     1381 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     1441 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     1501 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1561 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1621 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1681 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1741 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1801 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1861 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1921 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg
     1981 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     2041 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     2101 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     2161 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     2221 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     2281 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     2341 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     2401 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     2461 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     2521 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     2581 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     2641 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     2701 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     2761 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     2821 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     2881 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     2941 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     3001 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     3061 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     3121 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     3181 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     3241 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     3301 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     3361 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     3421 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     3481 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     3541 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
     3601 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     3661 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     3721 tttattacag ggacagcaga gatccagttt ggttaattaa agtgcaagtg ggttttagga
     3781 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
     3841 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
     3901 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
     3961 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
     4021 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
     4081 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
     4141 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
     4201 gagcgcagac cggtggtacc ataacttcgt ataggatact ttatacgaag ttatgcagaa
     4261 tggtagctgg attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca
     4321 tacattatac gaagttatac tacccaggct agcccggtgc caccatcacc ggatccggga
     4381 cctgaaataa aagacaaaaa gactaaactt acccaggcga atctgctgtt tcccctggcc
     4441 agagggccca cgtagtaggg gatgcggaag gtcaggatct tctcgatctt ttcccggttg
     4501 tccttcagga atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag ctctcccagg
     4561 tggatctggt gggggatgct gccgttgtcg aaggtccgct gcttccgcag caggtcctct
     4621 ctgttcagct tcacgagcag ttcctcggtg ccgtccatct tttccaggat gggcttgatg
     4681 aacttgtaga actcttcctg gctggctccg ccgtcaatgt agccggcgta gccgttcttg
     4741 ctctggtcga agaaaatctc tttgtacttc tcaggcagct gctgccgcac gagagctttc
     4801 agcagggtca ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc gctcaggggg
     4861 gccttggtga tctcggtgtt cactctcagg atgtcgctca gcaggatggc gtcggacagg
     4921 ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga tctgggccag caggttgtcc
     4981 aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc caggtcgaag
     5041 ttgctcttga agttgggggt caggcccagg ctcagggcaa tcaggttgcc gaacaggcca
     5101 ttcttcttct cgccgggcag ctgggcgatc agattttcca gccgtctgct cttgctcagt
     5161 ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt ttcctcgaac
     5221 agctggttgt aggtctgcac cagctggatg aacagcttgt ccacgtcgct gttgtcgggg
     5281 ttcaggtcgc cctcgatcag gaagtggccc cggaacttga tcatgtgggc cagggccaga
     5341 tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca gtttctttct caggtggtag
     5401 atggtggggt acttctcgtg gtaggccacc tcgtccacga tgttgccgaa gatggggtgc
     5461 cgctcgtgct tcttatcctc ttccaccagg aaggactctt ccagtctgtg gaagaagctg
     5521 tcgtccacct tggccatctc gttgctgaag atctcttgca gatagcagat ccggttcttc
     5581 cgtctggtgt atcttcttct ggcggttctc ttcagccggg tggcctcggc tgtttcgccg
     5641 ctgtcgaaca gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc ggtgttgccc
     5701 agcaccttga atttcttgct gggcaccttg tactcgtcgg tgatcacggc ccagcccaca
     5761 gagttggtgc cgatggccag gccgatgctg tacttcttgt cggctgctgg gactccgtgg
     5821 ataccgacct tccgcttctt ctttggggcc atcttatcgt catcgtcttt gtaatcaata
     5881 tcatgatcct tgtagtctcc gtcgtggtcc ttatagtcca tggtggcata acttcgtata
     5941 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     6001 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatgtggt gcaaatcaaa
     6061 gaactgctcc tcagtggatg ttgcctttac ttctagatga ccagaaagag cgaggaaacc
     6121 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     6181 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     6241 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     6301 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     6361 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     6421 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     6481 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     6541 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     6601 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     6661 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     6721 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     6781 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     6841 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     6901 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     6961 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     7021 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     7081 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     7141 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     7201 ctgtccgact acgatgtgga cgctatcgtg cctcagagct ttctgaagga cgactccatc
     7261 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     7321 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     7381 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     7441 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     7501 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     7561 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     7621 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     7681 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     7741 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     7801 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     7861 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     7921 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     7981 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     8041 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     8101 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     8161 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     8221 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     8281 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     8341 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     8401 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     8461 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     8521 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     8581 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     8641 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     8701 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     8761 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     8821 gccacgaaaa aggccggaca ggccaaaaag aaaaagctcg agggcggagg cgggagcgga
     8881 tccgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt
     8941 gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt
     9001 gacgcccttg atgatttcga cctggacatg ctgattaact ctagaagttc cggatctccg
     9061 aaaaagaaac gcaaagttgg tagccagtac ctgcccgaca ccgacgaccg gcaccggatc
     9121 gaggaaaagc ggaagcggac ctacgagaca ttcaagagca tcatgaagaa gtcccccttc
     9181 agcggcccca ccgaccctag acctccacct agaagaatcg ccgtgcccag cagatccagc
     9241 gccagcgtgc caaaacctgc cccccagcct taccccttca ccagcagcct gagcaccatc
     9301 aactacgacg agttccctac catggtgttc cccagcggcc agatctctca ggcctctgct
     9361 ctggctccag cccctcctca ggtgctgcct caggctcctg ctcctgcacc agctccagcc
     9421 atggtgtctg cactggctca ggcaccagca cccgtgcctg tgctggctcc tggacctcca
     9481 caggctgtgg ctccaccagc ccctaaacct acacaggccg gcgagggcac actgtctgaa
     9541 gctctgctgc agctgcagtt cgacgacgag gatctgggag ccctgctggg aaacagcacc
     9601 gatcctgccg tgttcaccga cctggccagc gtggacaaca gcgagttcca gcagctgctg
     9661 aaccagggca tccctgtggc ccctcacacc accgagccca tgctgatgga ataccccgag
     9721 gccatcaccc ggctcgtgac aggcgctcag aggcctcctg atccagctcc tgcccctctg
     9781 ggagcaccag gcctgcctaa tggactgctg tctggcgacg aggacttcag ctctatcgcc
     9841 gatatggatt tctcagcctt gctgggctct ggcagcggca gccgggattc cagggaaggg
     9901 atgtttttgc cgaagcctga ggccggctcc gctattagtg acgtgtttga gggccgcgag
     9961 gtgtgccagc caaaacgaat ccggccattt catcctccag gaagtccatg ggccaaccgc
    10021 ccactccccg ccagcctcgc accaacacca accggtccag tacatgagcc agtcgggtca
    10081 ctgaccccgg caccagtccc tcagccactg gatccagcgc ccgcagtgac tcccgaggcc
    10141 agtcacctgt tggaggatcc cgatgaagag acgagccagg ctgtcaaagc ccttcgggag
    10201 atggccgata ctgtgattcc ccagaaggaa gaggctgcaa tctgtggcca aatggacctt
    10261 tcccatccgc ccccaagggg ccatctggat gagctgacaa ccacacttga gtccatgacc
    10321 gaggatctga acctggactc acccctgacc ccggaattga acgagattct ggataccttc
    10381 ctgaacgacg agtgcctctt gcatgccatg catatcagca caggactgtc catcttcgac
    10441 acatctctgt tttgagaatt caattctaac tagacgcgtt aagtcgacaa tcaacctctg
    10501 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
    10561 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
    10621 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
    10681 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
    10741 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
    10801 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
    10861 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
    10921 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
    10981 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
    11041 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
    11101 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
    11161 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
    11221 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
    11281 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
    11341 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
    11401 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
    11461 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
    11521 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
    11581 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
    11641 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
    11701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
    11761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    11821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    11881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    11941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    12001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    12061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
    12121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
    12181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
    12241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
    12301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
    12361 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
    12421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
    12481 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
    12541 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
    12601 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
    12661 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
    12721 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
    12781 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
    12841 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
    12901 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
    12961 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
    13021 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
    13081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
    13141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
    13201 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
    13261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    13321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    13381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
    13441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    13501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    13561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    13621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    13681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    13741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    13801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    13861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    13921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    13981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    14041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
    14101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    14161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    14221 aaaaggatct caagaaga
//
