LOCUS       sequence-234973-        9898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor construct to amplify 
            NLS-Cas9m4-NLS-VP64-TaV-2a-Citrine-FRT-Kan-FRT cassette for BAC 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9898)
  AUTHORS   Chong-Morrison V, Mayes S, Simoes FC, Senanayake U, Carroll DS, 
            Riley PR, Wilson SW, Sauka-Spengler T
  TITLE     Ac/Ds transposition for CRISPR/dCas9-SID4x epigenome modulation in 
            zebrafish.
  JOURNAL   Biol Open. 2023 Jun 15;12(6):bio059995. doi: 10.1242/bio.059995. 
            Epub 2023 Jun 27.
  PUBMED    37367831
REFERENCE   2  (bases 1 to 9898)
  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"; journalName: "Biol 
            Open."; date: "2023-06-15"; volume: "12(6):bio059995. doi"; pages: "
            10.1242/bio.059995. Epub 2023 Jun 27"
COMMENT     Sequence Label: 
            pGEM-T-Easy-NLS-Cas9m4-NLS-VP64-TaV-2a-Citrine-FRT-Kan-FRT
FEATURES             Location/Qualifiers
     source          1..9898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..61
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     53..70
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     54..70
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     77..96
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        77..95
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      126..135
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      130..139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             148..168
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(178..204,205..207,208..2691,2692..2697,2698..2763,
                     2764..2766,2767..4281)
                     /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             4294..4314
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4339..4488
                     /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             4516..4569
                     /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(4576..4578,4579..5289)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4618..4639)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4879..4898)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5226..5247
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5344..5465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5355..5374)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5409..5428
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(5589..5636)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        5730..5834
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5836..6026
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5859..5879)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             6066..6860
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6066..6860
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6120..6139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6730..6749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6867..6915
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    complement(6940..6987)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(7116..7134)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7117..7134)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7152..7168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7152..7168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7165..7187)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7176..7192
                     /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(7200..7206,7207..7224,7225..7230))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7245..7266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7383..7400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7554..8142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7634..7653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8313..9104,9105..9173))
                     /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     8936..8955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9174..9278)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9356..9811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9493..9514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9705..9724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
       61 gacggccagt gaattgtaat acgactcact atagggcgaa ttgggcccga cgtcgcatgc
      121 tcccggccgc catggcggcc gcggatgccc aagaagaaga ggaaggtggg aagggggatg
      181 gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggctg ggccgtcatt
      241 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
      301 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
      361 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
      421 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
      481 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
      541 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
      601 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
      661 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
      721 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
      781 aatgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
      841 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
      901 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
      961 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     1021 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     1081 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     1141 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     1201 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     1261 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     1321 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     1381 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     1441 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     1501 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     1561 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     1621 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     1681 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     1741 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     1801 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     1861 aaacagctca aagaggacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     1921 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     1981 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     2041 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     2101 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     2161 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     2221 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     2281 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtctccac
     2341 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     2401 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     2461 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     2521 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     2581 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     2641 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggctgctatc
     2701 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     2761 aaagctagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     2821 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     2881 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     2941 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     3001 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     3061 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     3121 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     3181 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     3241 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     3301 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     3361 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     3421 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     3481 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     3541 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     3601 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     3661 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     3721 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     3781 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     3841 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     3901 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     3961 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     4021 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     4081 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     4141 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     4201 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     4261 ctctctcagc tcggtggaga cagcagggct gaccccaaga agaagaggaa ggtggaggcc
     4321 agcggttccg gacgggctga cgcattggac gattttgatc tggatatgct gggaagtgac
     4381 gccctcgatg attttgacct tgacatgctt ggttcggatg cccttgatga ctttgacctc
     4441 gacatgctcg gcagtgacgc ccttgatgat ttcgacctgg acatgctgat taactctaga
     4501 gatatcggat caggagaggg caggggaagt cttctaacat gcggggacgt ggaggaaaat
     4561 cccgggccca gatctgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     4621 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     4681 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     4741 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc
     4801 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     4861 cgcaccatct tcttcaaaga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     4921 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     4981 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     5041 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     5101 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     5161 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     5221 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     5281 ctgtacaagt aaagcggccg cgactctaga gtcggggcgg ccggccgctt cgagcagaca
     5341 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     5401 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     5461 aagttaacaa caacaattgc attcatttta tgtttcgggt tcagggggag gtgtgggagg
     5521 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg
     5581 acctgcttga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga
     5641 tccccctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     5701 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5761 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5821 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     5881 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5941 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6001 tctcaattag tcagtaacca tagtccgcaa agatcgatca agagacagga tgaggatcgt
     6061 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     6121 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6181 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6241 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6301 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6361 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6421 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6481 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6541 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     6601 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6661 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6721 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6781 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6841 ttcttgacga gttcttctga ggggatcggc aataaaaaga cagaataaaa cgcacgggtg
     6901 ttgggtcgtt tgttcggatc cgagcttcaa aagcgctctg aagttcctat actttctaga
     6961 gaataggaac ttcggaatag taacttctcc atggtagcct ccaaaaaagc ctcctcacta
     7021 cttctggaat agctcaatca ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga
     7081 gagctcccaa cgcgttggat gcatagcttg agtattctat agtgtcacct aaatagcttg
     7141 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7201 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7261 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7321 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7381 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7441 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7501 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7561 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7621 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7681 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7741 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7801 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7861 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7921 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7981 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8041 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8101 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8161 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8221 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8281 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8341 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8401 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8461 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8521 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8581 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8641 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8701 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8761 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8821 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8881 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8941 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     9001 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     9061 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9121 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9181 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9241 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9301 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     9361 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     9421 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     9481 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     9541 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     9601 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     9661 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     9721 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     9781 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     9841 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatg
//
