LOCUS       sequence-351670-       10173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cRaf fused to EGFP-eDHFR(69K6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10173)
  AUTHORS   Suzuki S, Nakamura A, Hatano Y, Yoshikawa M, Yoshii T, Sawada S, 
            Atsuta-Tsunoda K, Aoki K, Tsukiji S
  TITLE     A chemogenetic platform for controlling plasma membrane signaling 
            and synthetic signal oscillation.
  JOURNAL   Cell Chem Biol. 2022 Sep 15;29(9):1446-1464.e10. doi: 
            10.1016/j.chembiol.2022.06.005. Epub 2022 Jul 13.
  PUBMED    35835118
REFERENCE   2  (bases 1 to 10173)
  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"; doi: 
            "10.1016/j.chembiol.2022.06.005"; journalName: "Cell Chem Biol"; 
            date: "2022-09-15- 15"; volume: "29"; issue: "9"; pages: 
            "1446-1464.e10"
COMMENT     Sequence Label: pPBpuro-EGFP-eDHFR(69K6)-cRaf
FEATURES             Location/Qualifiers
     source          1..10173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          111..1127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1051..1073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1135..1154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1218..1227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1224..1226,1227..1229,1230..1940)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1224..1226,1227..1229,1230..1940)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1269..1290)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1530..1549)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1877..1898
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4586..5135
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4730..4747)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4957..4976
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5138..5737
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5138..5157)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5634..5654
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5764..5781)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5770..5994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6041..6057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6065..6081
                     /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(6089..6095,6096..6113,6114..6119))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6134..6155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6389..6451
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6492..6512)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6492..6510)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6531..6547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6555..6571
                     /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(6579..6585,6586..6603,6604..6609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6624..6645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6762..6779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6933..7521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7013..7032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7692..8483,8484..8552))
                     /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     8315..8334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8553..8657)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8683..9138)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8820..8841)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9032..9051
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9265..9287
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9279..9296
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9280..9296
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9306..9325
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9306..9324
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   9354..9388
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        9691..10070
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
       61 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      121 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      181 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      241 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      301 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      361 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      421 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
      481 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
      541 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
      601 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
      661 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
      721 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
      781 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
      841 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
      901 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
      961 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1021 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1081 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1141 tgctggttat tgtgctgtct catcattttg gcaaagaatt gggatcctca cgtcaattgt
     1201 agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1261 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1321 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1381 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1441 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1501 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1561 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1621 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1681 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1741 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1801 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1861 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1921 gcatggacga gctgtacaag tctgggtccg cagccagtag cgccggaagt atgatcagtc
     1981 tgattgcggc gttagcggta gatcgcgtta tcggcatgga aaacgccatg ccgtggaacc
     2041 tgcctgccga tctcgcctgg tttaaacgca acaccttaaa taaacccgtg attatgggcc
     2101 gccatacctg ggaatcaatc ggtcgtccgt tgccaggacg caaaaatatt atcctcagca
     2161 gtcaaccggg tacggacaaa aaaaagaaaa agaaagatcg cgtaacgtgg gtgaagtcgg
     2221 tggatgaagc catcgcggcg tgtggtgacg taccagaaat catggtgatt ggcggcggtc
     2281 gcgtttatga acagttcttg ccaaaagcgc aaaaactgta tctgacgcat atcgacgcag
     2341 aagtggaagg cgacacccat ttcccggatt acgagccgga tgactgggaa tcggtattca
     2401 gcgaattcca cgatgctgat gcgcagaact ctcacagcta ttgctttgag attctggagc
     2461 ggcggagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     2521 gtggtcctcg aagcctcgag atggagcaca tacagggagc ttggaagacg atcagcaatg
     2581 gttttggatt caaagatgcc gtgtttgatg gctccagctg catctctcct acaatagttc
     2641 agcagtttgg ctatcagcgc cgggcatcag atgatggcaa actcacagat ccttctaaga
     2701 caagcaacac tatccgtgtt ttcttgccga acaagcaaag aacagtggtc aatgtgcgaa
     2761 atggaatgag cttgcatgac tgccttatga aagcactcaa ggtgaggggc ctgcaaccag
     2821 agtgctgtgc agtgttcaga cttctccacg aacacaaagg taaaaaagca cgcttagatt
     2881 ggaatactga tgctgcgtct ttgattggag aagaacttca agtagatttc ctggatcatg
     2941 ttcccctcac aacacacaac tttgctcgga agacgttcct gaagcttgcc ttctgtgaca
     3001 tctgtcagaa attcctgctc aatggatttc gatgtcagac ttgtggctac aaatttcatg
     3061 agcactgtag caccaaagta cctactatgt gtgtggactg gagtaacatc agacaactct
     3121 tattgtttcc aaattccact attggtgata gtggagtccc agcactacct tctttgacta
     3181 tgcgtcgtat gcgagagtct gtttccagga tgcctgttag ttctcagcac agatattcta
     3241 cacctcacgc cttcaccttt aacacctcca gtccctcatc tgaaggttcc ctctcccaga
     3301 ggcagaggtc gacatccaca cctaatgtcc acatggtcag caccaccctg cctgtggaca
     3361 gcaggatgat tgaggatgca attcgaagtc acagcgaatc agcctcacct tcagccctgt
     3421 ccagtagccc caacaatctg agcccaacag gctggtcaca gccgaaaacc cccgtgccag
     3481 cacaaagaga gcgggcacca gtatctggga cccaggagaa aaacaaaatt aggcctcgtg
     3541 gacagagaga ttcaagctat tattgggaaa tagaagccag tgaagtgatg ctgtccactc
     3601 ggattgggtc aggctctttt ggaactgttt ataagggtaa atggcacgga gatgttgcag
     3661 taaagatcct aaaggttgtc gacccaaccc cagagcaatt ccaggccttc aggaatgagg
     3721 tggctgttct gcgcaaaaca cggcatgtga acattctgct tttcatgggg tacatgacaa
     3781 aggacaacct ggcaattgtg acccagtggt gcgagggcag cagcctctac aaacacctgc
     3841 atgtccagga gaccaagttt cagatgttcc agctaattga cattgcccgg cagacggctc
     3901 agggaatgga ctatttgcat gcaaagaaca tcatccatag agacatgaaa tccaacaata
     3961 tatttctcca tgaaggctta acagtgaaaa ttggagattt tggtttggca acagtaaagt
     4021 cacgctggag tggttctcag caggttgaac aacctactgg ctctgtcctc tggatggccc
     4081 cagaggtgat ccgaatgcag gataacaacc cattcagttt ccagtcggat gtctactcct
     4141 atggcatcgt attgtatgaa ctgatgacgg gggagcttcc ttattctcac atcaacaacc
     4201 gagatcagat catcttcatg gtgggccgag gatatgcctc cccagatctt agtaagctat
     4261 ataagaactg ccccaaagca atgaagaggc tggtagctga ctgtgtgaag aaagtaaagg
     4321 aagagaggcc tctttttccc cagatcctgt cttccattga gctgctccaa cactctctac
     4381 cgaagatcaa ccggagcgct tccgagccat ccttgcatcg ggcagcccac actgaggata
     4441 tcaatgcttg cacgctgacc acgtccccga ggctgcctgt cttctgcggc cgctagtcga
     4501 cggtaccgcg ggcccgggat aagtcaacta acttaagcta gcaacggttt ccctctagcg
     4561 ggatcaattc cgcccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     4621 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     4681 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     4741 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     4801 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     4861 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     4921 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     4981 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     5041 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     5101 ggggacgtgg ttttcctttg aaaaacacga taataccatg accgagtaca agcccacggt
     5161 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     5221 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     5281 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     5341 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5401 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5461 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5521 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5581 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5641 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5701 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatcg atgatctaga gctcgctgat
     5761 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5821 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5881 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5941 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     6001 aggcggaaag aacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6061 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6121 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6181 tccagtcggg aaacctgtcg tgccagcgga tcttaaaagt tttgttactt tatagaagaa
     6241 attttgagtt tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat
     6301 ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat
     6361 aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa
     6421 cgtacgtcac aatatgatta tctttctagg gttaatctag tatacgcgtt taattaatcc
     6481 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     6541 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6601 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6721 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7621 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7801 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7981 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8041 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8101 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8161 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8221 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8281 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8341 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8401 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8461 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8521 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8581 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8641 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     8701 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     8761 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     8821 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     8881 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     8941 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     9001 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     9061 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     9121 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9181 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9241 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     9301 gcgcgtaata cgactcacta tagggcgaat tggggcgcgc cattctagat taaccctaga
     9361 aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat attgctctct ctttctaaat
     9421 agcgcgaatc cgtcgctgtg catttaggac atctcagtcg ccgcttggag ctcccgtgag
     9481 gcgtgcttgt caatgcggta agtgtcactg attttgaact ataacgaccg cgtgagtcaa
     9541 aatgacgcat gattatcttt tacgtgactt ttaagattta actcatacga taattatatt
     9601 gttatttcat gttctactta cgtgataact tattatatat atattttctt gttatagata
     9661 tcaactagaa tgctagcatg ggcccatctc gacattgatt attgactagt tattaatagt
     9721 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     9781 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     9841 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     9901 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     9961 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
    10021 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
    10081 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
    10141 tatttatttt ttaattattt tgtgcagcga tgg
//
