LOCUS       sequence-496077-       14806 bp    DNA     circular SYN 02-JAN-2026
DEFINITION  Lentiviral expression of dCas9-VP64 with H2B-mCherry reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14806)
  TITLE     Brock Lab Plasmids 2025
REFERENCE   2  (bases 1 to 14806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 2, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: lenti-EF1a-dCas9-VP64-H2B-mCherry
FEATURES             Location/Qualifiers
     source          1..14806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      337..346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             343..1053
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(488..506)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     756..775
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1079..1667
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1132..1152)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1550..1561)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1670..1686)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1671..1687)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2192..2372
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2398..2415)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2404..2628
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2674..3102
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2761..2780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2971..2992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3111..3131)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3116..3445
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3296..3431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3358..3377
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3493..3540
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3559..3933
                     /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    4063..4184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4100..4119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4154..4173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4233..4249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4233..4249)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4246..4268)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4257..4273
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4281..4287,4288..4305,4306..4311))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4326..4347
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4464..4481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4635..5223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4715..4734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5394..6185,6186..6254))
                     /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     6017..6036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6255..6359)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6434..6453)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6625..7004
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7006..7204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7159..7179
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7222..7402
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7449..7574
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8067..8300
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8485..8529
                     /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    8827..8944
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        9074..10252
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9305..10243
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     10200..10220
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             10268..10315
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(10316..10339,10340..10342,10343..12898,12899..12901,
                     12902..14416)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             14471..14491
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14516..14665
                     /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             14687..14740
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(14753..14806,1..324)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
ORIGIN
        1 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
       61 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
      121 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
      181 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
      241 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
      301 atcaccaagt acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag
      361 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
      421 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
      481 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
      541 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
      601 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
      661 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
      721 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
      781 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
      841 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
      901 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
      961 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     1021 tccaccggcg gcatggacga gctgtacaag taattcgaac gatatcaagc ttatcggtaa
     1081 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1141 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1201 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     1261 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     1321 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     1381 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     1441 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     1501 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     1561 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     1621 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     1681 acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     1741 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     1801 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     1861 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     1921 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     1981 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     2041 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     2101 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     2161 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     2221 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     2281 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     2341 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
     2401 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2461 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2521 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2581 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2641 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2701 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2761 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2821 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2881 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2941 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3001 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3061 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3121 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3361 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3421 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3481 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     3541 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     3601 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     3661 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     3721 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     3781 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     3841 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3901 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     3961 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4021 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4081 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4141 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4201 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4261 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4321 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4381 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4441 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4501 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4561 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4621 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4681 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4741 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4801 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4861 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4921 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4981 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5041 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5101 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5161 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5221 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5281 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5341 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5401 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5461 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5521 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5581 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5641 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5701 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5761 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5821 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5881 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5941 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6001 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6061 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6121 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6181 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6241 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6301 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6361 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     6421 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     6481 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     6541 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     6601 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     6661 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6721 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6781 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6841 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     6901 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     6961 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     7021 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     7081 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     7141 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     7201 gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     7261 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7321 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7381 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     7441 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     7501 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     7561 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     7621 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7681 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7741 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7801 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7861 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7921 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     7981 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     8041 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     8101 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     8161 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     8221 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     8281 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     8341 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     8401 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     8461 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     8521 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     8581 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     8641 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     8701 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     8761 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat
     8821 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     8881 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     8941 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattagct agctgcaaag
     9001 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
     9061 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     9121 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     9181 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     9241 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     9301 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
     9361 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
     9421 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     9481 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     9541 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     9601 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     9661 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     9721 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     9781 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     9841 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     9901 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     9961 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
    10021 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
    10081 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
    10141 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
    10201 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gacgtacggc
    10261 caccatgaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggacaa
    10321 gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga
    10381 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
    10441 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
    10501 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
    10561 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
    10621 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
    10681 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
    10741 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
    10801 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
    10861 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
    10921 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
    10981 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc
    11041 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
    11101 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
    11161 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
    11221 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
    11281 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
    11341 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
    11401 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
    11461 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
    11521 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
    11581 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
    11641 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
    11701 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
    11761 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
    11821 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
    11881 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
    11941 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
    12001 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
    12061 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
    12121 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
    12181 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
    12241 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
    12301 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
    12361 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
    12421 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
    12481 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
    12541 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
    12601 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
    12661 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
    12721 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
    12781 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc acatcgtgcc
    12841 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaaggc
    12901 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
    12961 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
    13021 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
    13081 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
    13141 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
    13201 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
    13261 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
    13321 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
    13381 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
    13441 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
    13501 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
    13561 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
    13621 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
    13681 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
    13741 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
    13801 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
    13861 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
    13921 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
    13981 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
    14041 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
    14101 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
    14161 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
    14221 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
    14281 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
    14341 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
    14401 tcagctggga ggcgacagcg ctggaggagg tggaagcgga ggaggaggaa gcggaggagg
    14461 aggtagcgga cctaagaaaa agaggaaggt ggcggccgct ggatccggac gggctgacgc
    14521 attggacgat tttgatctgg atatgctggg aagtgacgcc ctcgatgatt ttgaccttga
    14581 catgcttggt tcggatgccc ttgatgactt tgacctcgac atgctcggca gtgacgccct
    14641 tgatgatttc gacctggaca tgctgattaa ctgtacaggt tcaggggagg gaagaggttc
    14701 cctgctcaca tgcggagacg tagaagagaa tcccggaccc atgaccttcg aaatgccaga
    14761 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcg
//
