LOCUS       sequence-460476-       14643 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A Tol2 based expression vector containing an Ubi driven loxP flanked
            GFP followed by an nls-mCherry reporter with fli1-TagBFP-zCre in the
            opposite direction. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14643)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda III J, Ortiz-Guzman J, Turner 
            WD, Largoza G, Sherman M, Mosser LE, Fujimoto E, Chien C-B, Kwan KM,
            Arenkiel BR, Devine WP, Wythe JD
  TITLE     Multisite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox with diverse applications in vertebrate model 
            systems
  JOURNAL   bioRxiv 2024.07.13.603267
REFERENCE   2  (bases 1 to 14643)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.07.13.603267"
COMMENT     Sequence Label: pTol2-Ubi-lsl-EGFP-nls-mCherry:fli1aep-CreI-zf...
FEATURES             Location/Qualifiers
     source          1..14643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1245..1265
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    complement(4788..4821)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4828..4837
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4834..4836,4837..4839,4840..5553)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4834..4836,4837..4839,4840..5550)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4879..4900)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5140..5159)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5487..5508
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5676..5797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6051..6084)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6115..6139
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6151..6171
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6172..6882
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGREFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6317..6335)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6585..6604
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6779..6802
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(6884..6908)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6910..7044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(7107..7127)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(7135..7151)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    7171..7252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    7776..7910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7932..7952)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7977..8675)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     intron          9501..9633
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(9868..9888)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      9891..9900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(12455..12475)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12455..12473)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12494..12510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12494..12510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12507..12529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12518..12534
                     /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(12542..12548,12549..12566,12567..12572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12587..12608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12725..12742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12896..13484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12976..12995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13655..14446,14447..14515))
                     /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     14278..14297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14516..14620)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctt
      661 aattaacaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta caagtactta
      721 gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact tgagtaaaag
      781 taaaaaagta ctccattaaa attgtacttg agtattaagg aagtaaaagt aaaagcaaga
      841 aagaaaacta gagattcttg tttaagctat ttaatctcaa aaaacattaa atgaaatgca
      901 tacaaggttt tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac
      961 aatctaatgc cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa
     1021 gctgattctc aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg
     1081 cagtgctgaa aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag
     1141 agaggctgca aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca
     1201 gatcctaggg ggcccgttta aacgccatga ttacgccaag ctatcaactt tgtatagaaa
     1261 agttggctcc gaattcgccc ttaaaactcg agaccagcaa agttctagaa tttgtcgaaa
     1321 catttatgtt atatatttcc tgaaaaaaat tctgagtaag ttcttaagtg ttattgccag
     1381 caacataaac aacagacggc aaaatgaata aatgataaca aagcagtagg cttaaataaa
     1441 cctaattttt ataggctgtt ctctacaacc ctcaaacagt gattagtttt gtacttataa
     1501 acttgccctt tcattcatat ttcaagaaaa ttggttcaga agatctggat attctagcag
     1561 ttgttcaagc tcatggaggg atcagtgacc tgattccaca atgactaggc ctaatccaga
     1621 aattagatga ctgtcaacat aaaaaggcac agcactcact agctgcccta tatattttat
     1681 tatattttac atatattatt ttatttattt agctctgagt gctgtacttt ctggttaaag
     1741 aaaactgctt acaacagcta acctgtacta cctcaggctc agggaatttg gaacaggttt
     1801 gtctggtttg tttctttaac catgcatgct tgttttcaac tatggcaaca cagtcacatg
     1861 ggacattaca gaaatgattt gtcgatgaca tgcgactttt ctttaataaa gcgcaaagat
     1921 cccaaaaagc aaacttttaa caaaaatcat ataattatat tttcaatcca gctttgtagc
     1981 aactttgtgc tgctgttcac tcagcaacag atagtcagta taaggtcagt gtgtctcaaa
     2041 gcagtgccat ctgtttcaca cattgcgttc tatatataag tgtgctggtt gacacgacac
     2101 tgtataaggc ctaggctaaa acacaaacaa tgtagaatga cactgtgttt tttttgtaaa
     2161 caaatgttgt ttttggttaa acatctttgt gaaaacatcc tcctgtcatg tatttgctat
     2221 attcaaatgt taaacccgtg cagaatagaa catatacaaa aaaaaacaac acaacacatt
     2281 tttaaacatt attaaatatc aagtattgct ggcagttctg tttctgtttt acagtaccct
     2341 ttgccacagt tctccgcttt tcctggtcca gattccacaa gtctgattca ccaatagcaa
     2401 agcgaataaa caaccaaagc agccaatcac tgcttgtaga ctgtcctgcg agaccggccc
     2461 attccagcac attctggaaa cttcctttat atgataatta taaatacatt taaattattg
     2521 atacaaaaca tgtaattcct agaacataac catagcaatc attagttttc agggtaatta
     2581 tgtattttta ggatttgact gcggaaagat ctggtcatgt gacgtctcat gaacgtcacg
     2641 gccctgggtt tctataaata cagtaggact ctcgaccatc ggcagatttt tcgaagaaga
     2701 agatcagttt caggagccgt actgttccgt ttcaacgcaa atattaacgg taagagcgaa
     2761 tttcctagtt tgttttcatg ccattcttta aaaccatagc gtattacttt aattatagta
     2821 aactttcgct ttctttatta caagagacgt tttgtgttga ttctccgcgg acattttcgg
     2881 tcagacaatc agaaaatgac cgcggaggac cagtaacttg cattacacgt aagttaaatc
     2941 ttcgtgtatt aaaatggtta ggttgttaac gtcaaatagg ttaccgtgtt tgcgtgtgat
     3001 caggttggtt ttgttagatt tttgtcagta tttttaattt atttgtttta gtttatttat
     3061 tttttttgct gaatcatagt ttgtgaacaa agaacccgga tgttacatac agtacagccg
     3121 ccatgttaca gagagttata acttaatcat tttaaaaata attttgcctt acttttagtt
     3181 tgtcatgttg agaaatgagg aaatgttaaa atgaggaaat atccaattaa tttaatatat
     3241 caaaataatc catgattaca atgcactgaa ctggagaaaa ttaagatgtt ttctagtgtc
     3301 atgaaacaaa tgtaagagat gtacattgta gatgttttat gtcaagaatt ggctagttga
     3361 tgcagcatac tggcgatact cagttgtaat aacagtaacg ttacatgtta atagactact
     3421 gagtatgctg ttctgtctat gtatgctctg taagctacga gaaggacttt tttaaacagt
     3481 aaagggtgca atatttttac aaattgaatt aaataaaggc tgtctattaa gtaatatgct
     3541 tgatattttt cttacttgat cggaaataag aaaaaatata aacgttgttg ctctaaaaat
     3601 cctagttcag tttagccaac cacaaatacc tttttgttcc tccaacagtt tttttttctt
     3661 ctctataata tttggcagtc tatagtactc caaatgtttc cccacagtct aactaattgg
     3721 tacagccaaa atcatgacac ttattgcaat aataattttg gttcattggc attgttgata
     3781 gcctgtgcca ctaatatggt cgattgatca tgcttcagga agaaaactat attgtttgat
     3841 gtaagattat taaatcttca cctgcttcca ttacaaacta ttcccatctt attgaattct
     3901 ggtatgtctt aaaggattag ttcacttccc aaatcaaaat ttacttagcc ttttttcatc
     3961 catgatccct ttttttcatc attaatgaag aaattgtttt tgaaaaagtt tcaagatttt
     4021 tttctctata ttgtggagct tgttagttta aaattccaaa atgcaatatg tggcttcaaa
     4081 tggttctaaa tgatcccagt caaggaataa cagtcttatc taatgaaacc attagacctt
     4141 taaaaataaa aaaaataaaa gtatttattt ttaaatgact gagtgattaa gttgaatttc
     4201 agcgtttcct tactgtgtag aagtccttcc ttactggccc caccctttgg ttctctgcca
     4261 atctgctacc taatgtaatg ttgtggaaca ttattattct ttatttctta attttttatt
     4321 ttttatttta aaacaatgta aactgcacag atgtgcagtt tgtttaaaat ggccaatgct
     4381 ttggaaatgc atgacataat tagatttcat gatgcacaaa gccaaatctc agagcttgtg
     4441 caaaatgagc tatcatttca ctaggtaaga ccctaaattt tcatatagga tcatttggac
     4501 aattttgctg caggtaaaat gcattctata gtccactgtc agccattgtt ttggatagta
     4561 tttatttttc tctacaagta tagtcaatag ttttctatta ttttaaaggt ttgtaacatt
     4621 taagggtgac caaatgcaaa gtaaaatttc attttcgggt gaactatctc gtttaacatg
     4681 ggagaagtgc aaaacataca ttattggcta gaacattgta gtatttttta aatggaaatg
     4741 tgtgattgct aatcttactt tgaatttgtt tacagggatc caagcttata acttcgtata
     4801 gcatacatta tacgaagtta tccggtcgcc accatggtga gcaagggcga ggagctgttc
     4861 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4921 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4981 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     5041 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     5101 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     5161 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     5221 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     5281 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     5341 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     5401 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     5461 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     5521 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     5581 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     5641 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     5701 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5761 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     5821 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5881 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5941 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6001 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ataacttcgt
     6061 atagcataca ttatacgaag ttataccggt ggatccttta agggcgaatt cgacccaagt
     6121 ttgtacaaaa aagcaggcta caccatggct ccaaagaaga agcgtaaggt aatggtgagc
     6181 aagggcgagg aggacaacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     6241 gagggctccg tgaacggccg cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     6301 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     6361 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     6421 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     6481 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     6541 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     6601 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     6661 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag
     6721 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     6781 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag
     6841 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagacccagc tttcttgtac
     6901 aaagtggggg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     6961 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     7021 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     7081 cagggggagg tgtgggaggt tttttccaac tttattatac atagttgata attcactggc
     7141 cgtcgtttta cggtaccatc gatgatgatc aacttgttta ttgcagctta taatggttac
     7201 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcgatatcaa
     7261 tcgggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
     7321 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
     7381 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
     7441 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaagctttag
     7501 gctgagtgca cggggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc
     7561 ccgctagggg gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca
     7621 tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct
     7681 ttcctctgaa cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa
     7741 aaagctttag gctgatacgt agtagatgcg gccgcaactt gtttattgca gcttataatg
     7801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gcgatcgcgt
     7921 ctagattatc atacctttct cttctttttt ggaccaccag caccacctcc tcgagcatta
     7981 agcttgtgcc ccagtttgct agggaggtcg cagtatctgg ccactgccac ctcgtgctgc
     8041 tcgacgtagg tctcgttgtt ggcctccttg attctttcca gtctgtagtc cacatagtag
     8101 acgccaggca tcttgaggtt cttagcgggt ttcttggatc tatatgtggt cttggcgttt
     8161 gcgatcagat ggctcccgcc cacgagcttc agggccatgt cgtttctgcc ttccaggccg
     8221 ccgtcagcgg ggtacagcgt ctcggtgaag gcctcccagc cgagtgtttt cttctgcatc
     8281 acagggccgt tggatgtgaa gttcacccct ctgatcttga cgttgtagat gaggcagccg
     8341 tcctggaggc tggtgtcctg ggtagcggtc agcacgcccc cgtcttcgta tgtggtgact
     8401 ctctcccatg tgaagccctc agggaaggac tgcttgaaga agtcggggat gccctgggtg
     8461 tggttgatga aggtcttgct gccgtagagg aagctagtag ccaggatgtc gaaggcgaag
     8521 gggagagggc cgccctcgac caccttgatt ctcatggtct gggtgccctc gtagggcttg
     8581 ccttcgccct cggatgtgca cttgaagtga tggttgtcca cggtgccctc catgtacagc
     8641 ttcatgtgca tgttctcctt aatcagctcg ctcatggtgg atccggcccc gccggctccg
     8701 ccgctagcgt ctccatcctc cagcagtctc accatagctc ctgtctcaga gtccaggttt
     8761 ctgatgtagt tcatcacaat gttcacattt gtccatcctc cagcctgcat gatctcaggg
     8821 atgctcactc cagctctagc catgtctcta gcagctccca ctctagcaga gtgtccagac
     8881 caagccaggt atctctgtcc agagtcatcc ttagctccat agatcagtct gtgtgtagcc
     8941 tcaaagattc cctccagagc ccttgtgctc agctggcttg tagcagaagg agcagccact
     9001 ccattctttc tcactctaca gaacaggtag ttgttagggt catcagccac tccagacaca
     9061 gagatccatc tctccaccag ctttgtcact cccaggctca gagccttctc cactccagct
     9121 gtgctcacca gtgtctttgt tcttccaatg tggatcagca ttcttcctcc atctgttctg
     9181 ctgatgtcct tcactctgat tctagcaatc tcagcaattc tcagcagtgt gttgtaagca
     9241 attcccagga aagccaggtt tctgatgtcc tgacatctgt cagagttctc catcaggctt
     9301 ctcacctggt caaagtctgt tctctcaaaa gccagagcct gcttagctct ctctccagca
     9361 tccacattct cctttctgat tcttctcatc accaggctca cagcattgct gtcagaaggt
     9421 ctaggcagtc cagaccttct gtgcagcatg ttcagctgtc ccaggtgctg ctggattgtc
     9481 ttcacagcca gtcctctagc ctgtggagag aaaggcaaag tggatgtcag taagaccaat
     9541 aggtgcctat cagaaacgca agagtcttct ctgtctcgac aagcccagtt tctattggtc
     9601 tccttaaacc tgtcttgtaa ccttgatact tacctggagg tacagcaggt agtctctcac
     9661 atcctcaggc tcagcaggga accactttct gttgttcagc ttacaccaag cagcccagct
     9721 cctgcagaca gacagcagca tcttccatgt gtgctcagag aaagcctgtc tgtctctgaa
     9781 catgtccatc aggttctttc tcacctcatc agatgtagca tccacaggca gagcaggtag
     9841 attttggtgg acggttagaa gattggagac tttcctcttc ttcttgggca ccatggtggc
     9901 catatgcggc acgcgtcgga tggttttttt tcctctaaat ttgggaagtg aagtcactgc
     9961 gcttgagaag gaagccgtgt atgcgtgtat gtgtgtgtgt gtatgtgtgt gtgtcactgg
    10021 ccctgccttc attacggcac ccaaagcagg atatttgtcc agcgaagata agacaccaaa
    10081 acaatggtgt ttctttcctt tgtgcatgtt tttgcctgga aatatttaca gaatgcctgc
    10141 caactcttta catttgttgc gctttttttc cgacattaaa caaggcgggt ttgtagaaac
    10201 attgcattga tgcatcaaag ctgtgctaga aataatcatt aatacaacgt ctgaaataac
    10261 agattatgct gattaatata ggctaaccat agtttaaggc taaaataaaa tatatacttt
    10321 ttcaacgatc gaatacaaat atgtctaatg cactcggtct aataataatc tcataatatg
    10381 atgaatattg ttccgttata tccaataact ttgatttgat ctacctatgg tgaagtgaag
    10441 aatgtgacga gtctgcccga gcttcttcta aaatgggctt ccatgaataa aaagcaaagc
    10501 ccgctgcgtc gagtctgtct gctccacacg tgtgcgcgcc tctgaatgca catgaacgca
    10561 gcgagcctgg tgctgtatgg accatataga gaggctgata cctcttgtgt gctgtgctgt
    10621 tcaacatttc tttaaatcag tagcatatga cctcacattt ttcatggaca ataataggaa
    10681 tattgtgcgt gttagactac atttactgtg cattaaatgt ctcatttcgt taaaaaaaga
    10741 aaaatagata tatttccgac ttgtagtgtt tagtgtcgaa acatattttg catgcattaa
    10801 atgcctgcta tttttatata gcatatacag cgtgcatgtg attccatctc taaatccaaa
    10861 atctccaccc cctctttttc agttaccgag cacgcattct tggtacattg aggtcgaaat
    10921 gctcacaaag ctgctatttt caacatggct ataaattggc cgtgtctgca aggctcgctt
    10981 aaaccatcca gaaccgcagg attacgctgg aatagagatt ttacgctgag gtgtgtcaag
    11041 agctcatctt tgtactgctg aatatgctgc atgatgaata atgcatgtat gctaccgtgc
    11101 ttttaactta acagggggac aaaaaactgt tttcataagc caacttgatc gtgattggct
    11161 gtcccgcaga agatgcgtca tgaggtcggg atgtacagag acccattaaa agcgctgtgc
    11221 tattttatac agagggcatt cctgtctttt gtttcaggag gaaaaggaaa cctgacagga
    11281 aaccagattg tgtggaacag gaaataccca tagattgaga cacccttctc attgtcgact
    11341 aaagtgtaca gttaaaagtc aatcatgtga ggtatctgtg tcaaattgtg aaggaaaaaa
    11401 catgcaggtt actattataa tctgtggatg catatgttta ctacactgaa aaacccataa
    11461 agttatggaa actcaaaccg tttgaggaaa ccgactgcaa caaaccattt aagttcaaaa
    11521 actaatccta atgagtactg tgaacttaat ctatttgatt aaacgatgca atttgagcac
    11581 atgttgaacc caataagtga agtgaactca aaccaactga gtactgtaaa acccaaaaag
    11641 ttaaggcaac tcaaactgct tgagggaacc gactgctaca aaccatttga gctaaactaa
    11701 tctatgtgag taccatgaac ttactccagt taagttgaag taatgaggta tttaatgaac
    11761 tcactacctt caacactgag tttaaaactc ttttcgaatg agtagaatca acattcagta
    11821 aattttgagt taattacact catttcattt gataaagttg actgttgggt tttagtgtat
    11881 gtttatgggt caagatgaga tactagtatc gatgcgaaga tacggccacg ggtgctcttg
    11941 atcctgtggc tgattttgga ctgtgctgct cgcagctgct gatgaatcac atacttcctc
    12001 cattttcttc cactgattga ctgttataat ttccctaatt tccaggtcaa ggtgctgtgc
    12061 attgtggtaa tagatgtgac atgacgtcac ttccaaagga ccaatgaaca tgtctgacca
    12121 atttcatata atgtgaaaac gattttcata ggcagaataa ataacattta aattaaactg
    12181 ggcatcagcg caattcaatt ggtttggtaa tagcaaggga aaatagaatg aagtgatctc
    12241 caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag tacttttttt
    12301 tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta ctcaagtaaa
    12361 gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac tcaagtactt
    12421 tacacctctg agatctgagc tccagctttt gttcccttta gtgagggtta attgcgcgct
    12481 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
    12541 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
    12601 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
    12661 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
    12721 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
    12781 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
    12841 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
    12901 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
    12961 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
    13021 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    13081 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    13141 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    13201 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    13261 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
    13321 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    13381 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    13441 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    13501 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
    13561 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
    13621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
    13681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
    13741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
    13801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
    13861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
    13921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
    13981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
    14041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
    14101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    14161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    14221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    14281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    14341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    14401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    14461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    14521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    14581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
    14641 cac
//
