LOCUS       sequence-379868-       15624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCas9-KRAB-MeCP2 with 2A Hygro 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15624)
  TITLE     Miner Lab plasmids
REFERENCE   2  (bases 1 to 15624)
  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: Lenti-EF1alpha-dCas9-KRAB-MeCP2_Hygro
FEATURES             Location/Qualifiers
     source          1..15624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    488..613
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             660..840
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(858..1056)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(883..903)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1058..1437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1609..1628
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1703..1807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1808..1876,1877..2668)
                     /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     complement(2026..2045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2839..3427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3328..3347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3581..3598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3715..3736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3751..3756,3757..3774,3775..3781)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3789..3805
                     /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)."
     primer_bind     3794..3816
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3813..3829
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3813..3829
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3878..3999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3889..3908)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3943..3962
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4129..4503)
                     /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"
     promoter        complement(4522..4569)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4617..4946)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4631..4766
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4685..4704)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4931..4951
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4960..5388)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5070..5091)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5282..5301
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5434..5658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5647..5664
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             5690..5870
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     6375..6391
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6376..6392
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6475..7653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6706..7644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7601..7621
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7666..7675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7672..7698,7699..7701,7702..10185,10186..10191,
                     10192..10257,10258..10260,10261..11775)
                     /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"
     CDS             11788..11808
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11869..12063
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             12097..12117
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12127..12984
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     CDS             12985..13038
                     /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             13042..14061
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     misc_feature    14101..14689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14154..14174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14572..14583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    14742..14859
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(15157..15201)
                     /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    15386..15619
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
       61 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
      121 tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta cttttgtttt
      181 gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca
      241 cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa
      301 gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag
      361 gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt
      421 ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct
      481 taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg
      541 gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc
      601 tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact
      661 gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca
      721 caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag
      781 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
      841 agtacaggca aaacgcgctg cttatataga cctcccaccg tacacgccta ccgcccattt
      901 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
      961 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     1021 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     1081 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     1141 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     1201 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     1261 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     1321 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     1381 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcaac
     1441 gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta agcagattct
     1501 tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact
     1561 cagcgacctc caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc
     1621 agattgtact gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg
     1681 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1741 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1801 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1861 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1921 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1981 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2041 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2101 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2161 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2221 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2281 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2341 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2401 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2461 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2521 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2581 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2641 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2701 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2761 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3541 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3601 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3661 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3721 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3781 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     3841 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
     3901 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3961 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
     4021 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     4081 caacctttca tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc
     4141 ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca
     4201 cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac
     4261 gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt
     4321 gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac
     4381 cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa
     4441 ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc
     4501 catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta
     4561 attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg
     4621 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag
     4681 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     4741 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     4801 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4861 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4921 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
     4981 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5041 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5101 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5161 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5221 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5281 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5341 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc
     5401 tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat
     5461 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     5521 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     5581 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     5641 ggcaactaga aggcacagtc gaggctgatc agcgggttta aacgggccct gctagagatt
     5701 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     5761 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     5821 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacagtcc
     5881 ggatgcagct ctcgggccat gtgatgaaat gctaggcggc tgtcaaacct ccactctaat
     5941 acttctctct ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gcgggtgttc
     6001 tctccttcat tggcttcttc taccttctct tgctcaactg gtactagctt gtagcaccat
     6061 ccaaaggtca gtggatatct gatccctggc cctggtgtgt agttctgcca atcagggaag
     6121 tagccttgtg tgtggtagat ccacagatca aggatatctt gtcttcgttg ggagtgaatt
     6181 agcccttcca gtcccccctt ttcttttaaa aagtggctaa gatctacagc tgccttgtaa
     6241 gtcattggtc ttaaaggtac ctgaggtgtg actggaaaac ccacctcctc ctcctcttgt
     6301 gcttctagcc aggcacaatc agcattggta gctgctgtat tgctacttgt gattgctcca
     6361 tgtttttcta ggtctcgagg tcgacggtat cgattgcaaa gatggataaa gttttaaaca
     6421 gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc
     6481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     6541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     6601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     6661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
     6721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
     6781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
     6841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
     6901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     6961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     7021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     7081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     7141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     7201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     7261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg aaggacgcgg
     7321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     7381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     7441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     7501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     7561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     7621 caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccgccac catggacaag
     7681 aagtactcca ttgggctcgc tatcggcaca aacagcgtcg gctgggccgt cattacggac
     7741 gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata
     7801 aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg
     7861 ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag
     7921 gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag
     7981 tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg
     8041 gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta
     8101 gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa
     8161 tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa
     8221 ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca
     8281 tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa
     8341 aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc
     8401 ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag
     8461 cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc
     8521 gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt
     8581 gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag
     8641 cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg
     8701 cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt
     8761 gacggcggag caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg
     8821 gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc
     8881 actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc
     8941 aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc
     9001 ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg
     9061 tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat
     9121 aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct
     9181 aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag
     9241 ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag
     9301 cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag
     9361 ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg
     9421 gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac
     9481 aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt
     9541 acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc
     9601 gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca
     9661 agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt
     9721 aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc
     9781 tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac
     9841 atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc
     9901 gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg
     9961 gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg
    10021 attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac
    10081 acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac
    10141 gtggatcagg aactggacat caatcggctc tccgactacg acgtggctgc tatcgtgccc
    10201 cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaagct
    10261 agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg
    10321 cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct
    10381 gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag
    10441 acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac
    10501 gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc
    10561 tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat
    10621 gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag
    10681 cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca
    10741 aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg
    10801 aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc
    10861 gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc
    10921 cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga
    10981 ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa
    11041 aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta
    11101 ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg
    11161 ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag
    11221 gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc
    11281 tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt
    11341 aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa
    11401 aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa
    11461 cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc
    11521 gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg
    11581 gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc
    11641 ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg
    11701 gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct
    11761 cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtgag tggtggagga
    11821 agtggcgggt cagggtcgat ggacgcgaaa tcacttacgg catggtcgag aacactggtt
    11881 acgttcaagg acgtgtttgt ggactttaca cgtgaggagt ggaaattgct ggatactgcg
    11941 caacaaattg tgtatcgaaa tgtcatgctt gagaattaca agaacctcgt cagtctcgga
    12001 taccagttga cgaaaccgga tgtgatcctt aggctcgaaa agggggaaga accttggctg
    12061 gtatcgggag gtggttcggg tggctctgga tcaagcccaa agaagaaacg gaaggtggaa
    12121 gcctcagtgc aggtgaaaag ggtgctggaa aaatcccccg gcaaactcct cgtgaagatg
    12181 cccttccagg cttcccctgg cggaaaaggt gaagggggtg gcgcaaccac atctgcccag
    12241 gtcatggtca tcaagcgacc tggaaggaaa agaaaggccg aggctgaccc tcaggccatt
    12301 ccaaagaaac ggggacgcaa gccagggtcc gtggtcgcag ctgcagcagc tgaggctaag
    12361 aaaaaggcag tgaaggaaag ctccatccgc agtgtgcagg agactgtcct gcccatcaag
    12421 aagaggaaga ctagggagac cgtgtccatc gaggtcaaag aagtggtcaa gcccctgctc
    12481 gtgtccaccc tgggcgaaaa atctggaaag gggctcaaaa catgcaagtc acctggacgg
    12541 aaaagcaagg agtctagtcc aaaggggcgc tcaagctccg cttctagtcc ccctaaaaag
    12601 gaacaccatc accatcacca tcacgccgag tctcctaagg ctcctatgcc actgctccca
    12661 ccacctccac cacctgagcc acagtcaagc gaagacccca tcagcccacc cgagcctcag
    12721 gatctgtcct ctagtatttg caaagaggaa aagatgccca gagcaggcag cctggagagt
    12781 gatggctgtc caaaagaacc cgccaagacc cagcctatgg tggcagccgc tgcaactacc
    12841 accacaacca caactaccac agtggccgaa aaatacaagc atcgcggcga gggcgaacga
    12901 aaggacattg tgtcaagctc catgcccaga cctaaccggg aggaaccagt cgatagtagg
    12961 acacccgtga ctgagagagt ctcagagggc agaggaagtc tgctaacatg cggtgacgtc
    13021 gaggagaatc ctggcccaac catgaaaaag cctgaactca ccgctacctc tgtcgagaag
    13081 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctccga gggcgaagaa
    13141 tctcgggctt tcagcttcga tgtgggaggg cgtggatatg tcctgcgggt gaatagctgc
    13201 gccgatggtt tctacaaaga tcgctatgtt tatcggcact ttgcatccgc cgctctccct
    13261 attcccgaag tgcttgacat tggggagttc agcgagagcc tgacctattg catctcccgc
    13321 cgtgcacagg gtgtcacctt gcaagacctg cctgaaaccg aactgcccgc tgttctccag
    13381 cccgtcgccg aggccatgga tgccatcgct gccgccgatc ttagccagac cagcgggttc
    13441 ggcccattcg gacctcaagg aatcggtcaa tacactacat ggcgcgattt catctgcgct
    13501 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcctcc
    13561 gtcgcccagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
    13621 ctcgtgcacg ccgatttcgg ctccaacaat gtcctgaccg acaatggccg cataacagcc
    13681 gtcattgact ggagcgaggc catgttcggg gattcccaat acgaggtcgc caacatcttc
    13741 ttctggaggc cctggttggc ttgtatggag cagcagaccc gctacttcga gcggaggcat
    13801 cccgagcttg caggatctcc tcggctccgg gcttatatgc tccgcattgg tcttgaccaa
    13861 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggctca gggtcgctgc
    13921 gacgcaatcg tccggtccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
    13981 gctgccgtct ggaccgatgg ctgtgtggaa gtgctcgccg atagtggaaa cagacgcccc
    14041 agcactcgtc ctagggcaaa ggatctgcag taatgagaat tcgatatcaa gcttatcggt
    14101 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
    14161 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
    14221 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
    14281 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
    14341 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
    14401 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
    14461 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
    14521 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
    14581 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
    14641 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgctagcta
    14701 attaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta
    14761 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
    14821 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
    14881 tgtctgcaga attggcgcac gcagtgccga tccgttcact aatcgaatgg atctgtctct
    14941 gtctctctct ccaccttctt cttctattcc ttcgggcctg tcgggtcccc tcggggttgg
    15001 gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag
    15061 tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga ataattttat
    15121 ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc
    15181 caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat
    15241 taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat
    15301 ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag
    15361 ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc
    15421 caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg
    15481 cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct
    15541 ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa
    15601 cccaaggaac aaagctccta ttcc
//
