LOCUS       sequence-354473-       15679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of dCas9-VPR-mCherry fusion protein for 
            CRISPRa..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15679)
  TITLE     Corn Lab Cas9 plasmids
REFERENCE   2  (bases 1 to 15679)
  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"
COMMENT     Sequence Label: IGI-P0492 pHR-dCas9-NLS-VPR-mCherry
FEATURES             Location/Qualifiers
     source          1..15679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1998..2016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2176..2195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2409..2428
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2718..2736
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2718..2735
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     LTR             2761..2994
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     3084..3100
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3085..3101
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3104..3692
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3210..3221
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3619..3639
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3750..4457)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4028..4047)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4297..4315
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4458..4514)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(4515..5084)
                     /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"
     CDS             complement(5103..5459)
                     /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             complement(5889..5909)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5934..6083)
                     /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"
     protein_bind    complement(6113..6137)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(6174..6194)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(6207..7721,7722..7724,7725..7790,7791..7796,
                     7797..10280,10281..10283,10284..10310))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     regulatory      10307..10316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(10404..10428)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(10425..10628)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(10454..10474)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        10629..11007
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    11068..11185
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(11470..11514)
                     /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    11699..11932
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    12429..12554
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             12601..13234
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    13402..13536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13426..13445)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     13480..13499
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(13961..13981)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          14111..14176
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        complement(15310..15639)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      15324..15459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     15624..15644
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct
     2221 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     2281 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     2341 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     2401 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     2461 ccggccacga tgcgtccggc gtagaggatc tggctagcga tgaccctgct gattggttcg
     2521 ctgaccattt ccgggtgcgg gacggcgtta ccagaaactc agaaggttcg tccaaccaaa
     2581 ccgactctga cggcagttta cgagagagat gatagggtct gcttcagtaa gccagatgct
     2641 acacaattag gcttgtacat attgtcgtta gaacgcggct acaattaata cataacctta
     2701 tgtatcatac acatacgatt taggtgacac tatagaatac acggaattaa ttctagatgc
     2761 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     2821 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     2881 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     2941 cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc
     3001 ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag
     3061 gtaccgagct cgaattaatt ccctcgaggt cgacggtatc gatgcgggga ggcggcccaa
     3121 agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc
     3181 agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac
     3241 gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc aaggaaagga cgatgatttc
     3301 cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca
     3361 aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa
     3421 aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg
     3481 cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag
     3541 gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc
     3601 attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat
     3661 ctttcacaaa ttttgtaatc cagaggttga ttatcgataa gcttgatatc aagcttgcat
     3721 gcctgcaggt cgactctaga gtcgcggccc tacttgtaca gctcgtccat gccgccggtg
     3781 gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag
     3841 gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc
     3901 ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc
     3961 ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag
     4021 cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc
     4081 accttgtaga tgaactcgcc gtcctggagg gaggagtcct gggtcacggt caccacgccg
     4141 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag
     4201 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg
     4261 gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc
     4321 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     4381 acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc
     4441 tcctcgccct tgctcacagg tccagggttc tcctccacgt ctccagcctg cttcagcagg
     4501 ctgaagttag tagcaaacag agatgtgtcg aagatggaca gtcctgtgct gatatgcatg
     4561 gcatgcaaga ggcactcgtc gttcaggaag gtatccagaa tctcgttcaa ttccggggtc
     4621 aggggtgagt ccaggttcag atcctcggtc atggactcaa gtgtggttgt cagctcatcc
     4681 agatggcccc ttgggggcgg atgggaaagg tccatttggc cacagattgc agcctcttcc
     4741 ttctggggaa tcacagtatc ggccatctcc cgaagggctt tgacagcctg gctcgtctct
     4801 tcatcgggat cctccaacag gtgactggcc tcgggagtca ctgcgggcgc tggatccagt
     4861 ggctgaggga ctggtgccgg ggtcagtgac ccgactggct catgtactgg accggttggt
     4921 gttggtgcga ggctggcggg gagtgggcgg ttggcccatg gacttcctgg aggatgaaat
     4981 ggccggattc gttttggctg gcacacctcg cggccctcaa acacgtcact aatagcggag
     5041 ccggcctcag gcttcggcaa aaacatccct tccctggaat cccggctgcc gctgccagag
     5101 cccagcaagg ctgagaaatc catatcggcg atagagctga agtcctcgtc gccagacagc
     5161 agtccattag gcaggcctgg tgctcccaga ggggcaggag ctggatcagg aggcctctga
     5221 gcgcctgtca cgagccgggt gatggcctcg gggtattcca tcagcatggg ctcggtggtg
     5281 tgaggggcca cagggatgcc ctggttcagc agctgctgga actcgctgtt gtccacgctg
     5341 gccaggtcgg tgaacacggc aggatcggtg ctgtttccca gcagggctcc cagatcctcg
     5401 tcgtcgaact gcagctgcag cagagcttca gacagtgtgc cctcgccggc ctgtgtaggt
     5461 ttaggggctg gtggagccac agcctgtgga ggtccaggag ccagcacagg cacgggtgct
     5521 ggtgcctgag ccagtgcaga caccatggct ggagctggtg caggagcagg agcctgaggc
     5581 agcacctgag gaggggctgg agccagagca gaggcctgag agatctggcc gctggggaac
     5641 accatggtag ggaactcgtc gtagttgatg gtgctcaggc tgctggtgaa ggggtaaggc
     5701 tggggggcag gttttggcac gctggcgctg gatctgctgg gcacggcgat tcttctaggt
     5761 ggaggtctag ggtcggtggg gccgctgaag ggggacttct tcatgatgct cttgaatgtc
     5821 tcgtaggtcc gcttccgctt ttcctcgatc cggtgccggt cgtcggtgtc gggcaggtac
     5881 tggctaccaa ctttgcgttt ctttttcgga gatccggaac ttctagagtt aatcagcatg
     5941 tccaggtcga aatcatcaag ggcgtcactg ccgagcatgt cgaggtcaaa gtcatcaagg
     6001 gcatccgaac caagcatgtc aaggtcaaaa tcatcgaggg cgtcacttcc cagcatatcc
     6061 agatcaaaat cgtccaatgc gtcagcccgt ccggaaccgc tggcctcttt gtagcctgct
     6121 tttttgtaca aacttgttga tatcaacgcg tcaagtcgac ggatccctgg cgacaccttc
     6181 ctcttcttct tggggtcagc cctgctgtct ccaccgagct gagagaggtc gattcttgtt
     6241 tcatagagcc ccgtaattga ctgatgaatc agtgtggcgt ccaggacctc ctttgtagag
     6301 gtgtaccgct ttctgtctat ggtggtgtcg aagtacttga aggctgcagg cgcgcccaag
     6361 ttggtcagag taaacaagtg gataatgttt tctgcctgct ccctgatggg cttatccctg
     6421 tgcttattgt aagcagaaag caccttatcg aggttagcgt cggcgaggat cactcttttg
     6481 gagaattcgc ttatttgctc gatgatctca tcaaggtagt gtttgtgttg ttccacgaac
     6541 agctgcttct gctcattatc ttcgggagac cctttgagct tttcatagtg gctggccaga
     6601 tacaagaaat taacgtattt agagggcagt gccagctcgt tacctttctg cagctcgccc
     6661 gcactagcga gcattcgttt ccggccgttt tcaagctcaa agagagagta cttgggaagc
     6721 ttaatgatga ggtctttttt gacctcttta tatcctttcg cctcgagaaa gtcgatgggg
     6781 tttttttcga agcttgatcg ctccatgatt gtgatgccca gcagttcctt gacgcttttg
     6841 agttttttag acttcccttt ctccactttg gccacaacca gtacactgta agcgactgta
     6901 ggagaatcga atccgccgta tttcttgggg tcccaatctt ttttgcgtgc gatcagcttg
     6961 tcgctgttcc ttttcgggag gatactttcc ttggagaagc ctccggtctg tacttcggtc
     7021 tttttaacga tgttcacctg cggcatggac aggaccttcc ggactgtcgc gaaatcccta
     7081 cccttgtccc acacgatttc tcctgtttct ccgtttgttt cgataagtgg tcgcttccga
     7141 atctctccat tggccagtgt aatctcggtc ttgaaaaaat tcataatatt gctgtaaaag
     7201 aagtacttag cggtggcctt gcctatttcc tgctcagact ttgcgatcat tttcctaaca
     7261 tcgtacactt tatagtctcc gtaaacaaat tcagattcaa gcttgggata ttttttgata
     7321 agtgcagtgc ctaccactgc attcaggtag gcatcatgcg catggtggta attgttgatc
     7381 tctctcacct tataaaactg aaagtccttt ctgaaatctg agaccagctt agacttcaga
     7441 gtaataactt tcacctctcg aatcagtttg tcattttcat cgtacttggt gttcatgcgt
     7501 gaatcgagaa tttgggccac gtgcttggtg atctggcgtg tctcaacaag ctgccttttg
     7561 atgaagccgg ctttatccaa ctcagacagg ccacctcgtt cagccttagt cagattatcg
     7621 aacttccgtt gtgtgatcag tttggcgttc agcagctgcc gccaataatt tttcattttc
     7681 ttgacaactt cttctgaggg gacgttatca ctcttccctc tagctttatc ggatcttgtc
     7741 aacactttat tatcaataga atcatctttg agaaaagact ggggcacgat agcagccacg
     7801 tcgtagtcgg agagccgatt gatgtccagt tcctgatcca cgtacatgtc cctgccgttc
     7861 tgcaggtagt acaggtagag cttctcattc tgaagctggg tgttttcaac tgggtgttcc
     7921 ttaaggattt gggaccccag ttcttttata ccctcttcaa tcctcttcat cctttcccta
     7981 ctgttcttct gtcccttctg ggtagtttgg ttctctcggg ccatctcgat aacgatattc
     8041 tcgggcttat gccttcccat tactttgacg agttcatcca cgaccttaac ggtctgcagt
     8101 attccctttt tgatagctgg gctacctgca agattagcga tgtgctcgtg aagactgtcc
     8161 ccctggccag aaacttgtgc tttctggatg tcctccttaa aggtgagaga gtcatcatgg
     8221 atcaactgca tgaagttccg gttggcaaat ccatcggact taagaaaatc caggattgtc
     8281 tttccactct gcttgtctcg gatcccattg atcagttttc ttgacagccg cccccatcct
     8341 gtatatcggc gcctcttgag ctgtttcatg actttgtcgt cgaagagatg agcgtaagtt
     8401 ttcaagcgtt cttcaatcat ctccctatct tcaaacaacg taagggtgag gacaatgtcc
     8461 tcaagaatgt cctcgttctc ctcattgtcc aggaagtcct tgtctttaat gattttcagg
     8521 agatcgtgat acgttcccag ggatgcgttg aagcgatcct ccactccgct gatttcaaca
     8581 gagtcgaaac attcaatctt tttgaaatag tcttctttga gctgtttcac ggtaactttc
     8641 cggttcgtct tgaagaggag gtccacgata gctttcttct gctctccaga caggaatgct
     8701 ggctttctca tcccttctgt gacgtatttg accttggtga gctcgttata aactgtgaag
     8761 tactcgtaca gcagagagtg tttaggaagc accttttcgt taggcagatt tttatcaaag
     8821 ttagtcatcc tttcgatgaa ggactgggca gaggccccct tatccacgac ttcctcgaag
     8881 ttccagggag tgatggtctc ttctgatttg cgagtcatcc acgcgaatct ggaatttccc
     8941 cgggcgaggg ggcctacata gtagggtatc cgaaatgtga ggattttctc aatcttttcc
     9001 ctgttatctt tcaaaaaggg gtagaaatcc tcttgccgcc tgaggatagc gtgcagttcg
     9061 cccaggtgaa tctggtgggg gatgcttcca ttgtcgaaag tgcgctgttt gcgcaacaga
     9121 tcttctctgt taagctttac cagcagctcc tcggtgccgt ccattttttc caagatgggc
     9181 ttaataaatt tgtaaaattc ctcctggctt gctccgccgt caatgtatcc ggcgtagcca
     9241 tttttagact gatcgaagaa aatttccttg tacttctcag gcagttgctg tctgacaagg
     9301 gccttcagca aagtcaagtc ttggtggtgc tcatcatagc gcttgatcat actagcgctc
     9361 agcggagctt tggtgatctc cgtgttcact cgcagaatat cactcagcag aatggcgtct
     9421 gacaggttct ttgccgccaa aaaaaggtct gcgtactggt cgccgatctg ggccagcaga
     9481 ttgtcgagat catcatcgta ggtgtctttg ctcagttgaa gcttggcatc ttcggccagg
     9541 tcgaagttag atttaaagtt gggggtcagc ccgagtgaca gggcgataag attaccaaac
     9601 aggccgttct tcttctcccc agggagctgt gcgatgaggt tttcgagccg ccgggatttg
     9661 gacagcctag cgctcaggat tgctttggcg tcaactccgg atgcgttgat cgggttctct
     9721 tcgaaaagct gattgtaagt ctgaaccagt tggataaaga gtttgtcgac atcgctgttg
     9781 tctgggttca ggtccccctc gatgaggaag tgtccccgaa atttgatcat atgcgccagc
     9841 gcgagataga tcaaccgcaa gtcagcctta tcagtactgt ctacaagctt cttcctcaga
     9901 tgatatatgg ttgggtactt ttcatggtac gccacctcgt ccacgatatt gccaaagatt
     9961 gggtggcgct cgtgcttttt atcctcctcc accaaaaagg actcctccag cctatggaag
    10021 aaagagtcat ccaccttagc catctcatta ctaaagatct cctgcaggta gcagatccga
    10081 ttctttctgc gggtatatct gcgccgtgct gttcttttga gccgcgtggc ttcggccgtc
    10141 tccccggagt cgaacaggag ggcgccaatg aggttcttct ttatgctgtg gcgatcggta
    10201 ttgcccagaa ctttgaattt tttgctcggc accttgtact cgtccgtaat gacggcccag
    10261 ccgacgctgt ttgtgccgat agcgagccca atggagtact tcttgtccat ggtggcaagg
    10321 gttcgatcct ctagagtccg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa
    10381 cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag
    10441 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
    10501 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
    10561 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
    10621 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
    10681 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
    10741 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
    10801 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
    10861 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
    10921 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
    10981 tattaataac tagtcaataa tcaatgtgaa ttcgatatca agcttatcga tccaaactgg
    11041 atctctgctg tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa
    11101 ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc
    11161 cccaatcccc ccttttcttt taaaattgtg gatgaatact gccatttggc gataccgtcg
    11221 agatccgttc actaatcgaa tggatctgtc tctgtctctc tctccacctt cttcttctat
    11281 tccttcgggc ctgtcgggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga
    11341 atatccctgc ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac
    11401 caagcctcct actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa
    11461 ccaattccac aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg
    11521 ctggttttgc gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct
    11581 gtcccactcc atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact
    11641 agcattccaa ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag
    11701 gagctgttga tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat
    11761 gccccagact gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt
    11821 ctgctgctgc actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc
    11881 tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac
    11941 tgctcttttt tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg
    12001 ttcaattttt actactttat atttatataa ttcacttctc caattgtccc tcatatctcc
    12061 tcctccaggt ctgaagatca gcggccggcc gcttgctgtg cggtggtctt acttttgttt
    12121 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
    12181 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
    12241 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
    12301 ggattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
    12361 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
    12421 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
    12481 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
    12541 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
    12601 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
    12661 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
    12721 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
    12781 cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca ctccccagtc
    12841 ccgcccaggc cacgcctccc tggaaagtcc ccagcggaaa gtcccttgta gcaagctcga
    12901 tatcagcagt tcttgaagta ctccggatgc agctctcggg ccacgtgatg aaatgctagg
    12961 cggctgtcaa acctccactc taacacttct ctctccgggt catccatccc atgcaggctc
    13021 acagggtgta acaagctggt gttctctcct ttattggcct cttctacctt atctggctca
    13081 actggtacta gcttgtagca ccatccaaag gtcagtggat atctgacccc tggccctggt
    13141 gtgtagttct gctaatcagg gaagtagcct tgtgtgtggt agatccacag atcaaggata
    13201 tcttgtcttc tttgggagtg aattagccct tccaactact aagtttgtag tacatattta
    13261 acaaatacaa tttctttaaa atgaaaataa ttcagaggaa tcacaggttt agagtaaatg
    13321 aaaccacagg taattggcag tggtaatagg gtatggggtg ggaagtttgg gatgattttg
    13381 gttagcttga gttatccagt tgatccagac atgataagat acattgatga gtttggacaa
    13441 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
    13501 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
    13561 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
    13621 tgtggtatgg ctgattatga tctctagtca aggcactata catcaaatat tccttattaa
    13681 cccctttaca aattaaaaag ctaaaggtac acaatttttg agcatagtta ttaatagcag
    13741 acactctatg cctgtgtgga gtaagaaaaa acagtatgtt atgattataa ctgttatgcc
    13801 tacttataaa ggttacagaa tatttttcca taattttctt gtatagcagt gcagcttttt
    13861 cctttgtggt gtaaatagca aagcaagcaa gagttctatt actaaacaca gcatgactca
    13921 aaaaacttag caattctgaa ggaaagtcct tggggtcttc tacctttctc ttcttttttg
    13981 gaggagtaga atgttgagag tcagcagtag cctcatcatc actagatggc atttcttctg
    14041 agcaaaacag gttttcctca ttaaaggcat tccaccactg ctcccattca tcagttccat
    14101 aggttggaat ctaaaataca caaacaatta gaatcagtag tttaacacat tatacactta
    14161 aaaattttat atttacctta gagctttaaa tctctgtagg tagtttgtcc aattatgtca
    14221 caccacagaa gtaaggttcc ttcacaaaga tccggggccc actcataaat ccagttgccg
    14281 ccacggtagc caatcaccgt atcgtataaa tcatcgtcgg tacgttcggc atcgctcatc
    14341 acaatacgtg cctggacgtc gaggatttcg cgtgggtcaa tgccgcgcca gatccacatc
    14401 agacggttaa tcatgcgata ccagtgaggg atggttttac catcaagggc cgactgcaca
    14461 ggcggttgtg cgccgtgatt aaagcggcgg actagcgtcg aggtttcagg atgtttaaag
    14521 cggggtttga acagggtttc gctcaggttt gcctgtgtca tggatgcagc ctccagaata
    14581 cttactggaa actattgtaa cccgcctgaa gttaaaaaga acaacgcccg gcagtgccag
    14641 gcgttgaaaa gattagcgac cggagattgg cgggacgaat acgacgccca tatcccacgg
    14701 ctgttcaatc caggtatctt gcgggatatc aacaacatag tcatcaacca gcggacgacc
    14761 agccggtttt gcgaagatgg tgacaaagtg cgcttttgga tacatttcac gaatcgcaac
    14821 cgcagtacca ccggtatcca ccaggtcatc aataacgatg aagccttcgc catcgccttc
    14881 tgcgcgtttc agcactttaa gctcgcgctg gttgtcgtga tcgtagctgg aaatacaaac
    14941 ggtatcgaca tgacgaatac ccagttcacg cgccagtaac gcacccggta cagaccgcca
    15001 cggcttacgg caataatgcc tttccattgt tcagaaggca tcagtcggct tgcgagttta
    15061 cgtgcatgga tctgcaacat gtcccaggtg acgatgtatt tttcgctcat gtgaagtgtc
    15121 ccagcctgtt tatctacggc ttaaaaagtg ttcgagggga aaataggttg cgcgagatta
    15181 tagagatctg gcgcactaaa aaccagtatt tcacatgagt ccgcgtcttt ttacgcactg
    15241 cctctccctg acgcgggata aagtggtatt ctcaaacata tctcgcaagc ctgtcttgtg
    15301 tccaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
    15361 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
    15421 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
    15481 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
    15541 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
    15601 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct gcattaatga
    15661 atcggccaac gcgcgggga
//
