LOCUS       sequence-340530-       12447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of RasGRF1(Cdc25) fused to 
            mNeonGreen-eDHFR(69K6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12447)
  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 12447)
  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: pCSIIpuro-mNG-eDHFR(69K6)-RasGEF
FEATURES             Location/Qualifiers
     source          1..12447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    85..202
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        258..1420
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          489..1411
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1368..1388
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1442..1451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1448..2155
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1703..1712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3629..4178
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3773..3790)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4000..4019
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4181..4780
                     /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(4181..4200)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4677..4697
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4815..5403
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4868..4888)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5286..5297)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5922..6102
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6128..6145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6134..6358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6404..6832
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6491..6510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6701..6722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6841..6861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6846..7175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7026..7161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7088..7107
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7223..7270
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7289..7663
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7793..7914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7830..7849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7884..7903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7963..7979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7963..7979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7976..7998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7987..8003
                     /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(8011..8017,8018..8035,8036..8041))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8056..8077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8194..8211)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8365..8953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8445..8464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9124..9915,9916..9984))
                     /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     9747..9766
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9985..10089)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10164..10183)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10355..10734
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10736..10934
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10889..10909
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10952..11132
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11179..11304
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11796..12029
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12214..12258
                     /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"
ORIGIN
        1 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
       61 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
      121 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
      181 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagaaatt cactttgaat
      241 taattcaagc ttcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      301 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      361 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      421 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      481 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      541 cttgcgtgcc ttgaattact tccacctggc tccagtacgt gattcttgat cccgagctgg
      601 agccaggggc gggccttgcg ctttaggagc cccttcgcct cgtgcttgag ttgaggcctg
      661 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      721 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      781 gcaagatagt cttgtaaatg cgggccagga tctgcacact ggtatttcgg tttttggggc
      841 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      901 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      961 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1021 agttgcgtga gcggaaagat ggccgcttcc cggccctgct ccagggggct caaaatggag
     1081 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaag gggcctttcc
     1141 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1201 ttagttctgg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1261 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1321 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1381 agtggttcaa agtttttttc ttccatttca ggtgtcgtga acacgctacc ggtccgaatt
     1441 ggccaccatg gtgagcaagg gcgaggagga taacatggcc tctctcccag cgacacatga
     1501 gttacacatc tttggctcca tcaacggtgt ggactttgac atggtgggtc agggcaccgg
     1561 caatccaaat gatggttatg aggagttaaa cctgaagtcc accaagggtg acctccagtt
     1621 ctccccctgg attctggtcc ctcatatcgg gtatggcttc catcagtacc tgccctaccc
     1681 tgacgggatg tcgcctttcc aggccgccat ggtagatggc tccggctacc aagtccatcg
     1741 cacaatgcag tttgaagatg gtgcctccct tactgttaac taccgctaca cctacgaggg
     1801 aagccacatc aaaggagagg cccaggtgaa ggggactggt ttccctgctg acggtcctgt
     1861 gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg aagaagactt accccaacga
     1921 caaaaccatc atcagtacct ttaagtggag ttacaccact ggaaatggca agcgctaccg
     1981 gagcactgcg cggaccacct acacctttgc caagccaatg gcggctaact atctgaagaa
     2041 ccagccgatg tacgtgttcc gtaagacgga gctcaagcac tccaagaccg agctcaactt
     2101 caaggagtgg caaaaggcct ttaccgatgt gatgggcatg gacgagctct acaagtctgg
     2161 gtccgcagcc agtagcgccg gaagtatgat cagtctgatt gcggcgttag cggtagatcg
     2221 cgttatcggc atggaaaacg ccatgccgtg gaacctgcct gccgatctcg cctggtttaa
     2281 acgcaacacc ttaaataaac ccgtgattat gggccgccat acctgggaat caatcggtcg
     2341 tccgttgcca ggacgcaaaa atattatcct cagcagtcaa ccgggtacgg acaaaaaaaa
     2401 gaaaaagaaa gatcgcgtaa cgtgggtgaa gtcggtggat gaagccatcg cggcgtgtgg
     2461 tgacgtacca gaaatcatgg tgattggcgg cggtcgcgtt tatgaacagt tcttgccaaa
     2521 agcgcaaaaa ctgtatctga cgcatatcga cgcagaagtg gaaggcgaca cccatttccc
     2581 ggattacgag ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca
     2641 gaactctcac agctattgct ttgagattct ggagcggcgg ctgtacaagt ccggactcag
     2701 atctcgacaa ggtagtggtg ctggctctgg tgctggtagt ggcgctggtt ccggtgctgg
     2761 ctctggcgcg cctcgagctc aagcttcgaa ttctacgctg gaggagatca cgcagatggc
     2821 tgaaggcgtg aaggctgagc cctttgaaaa ccactcagcc ctggagatcg cggagcagct
     2881 gaccctgcta gatcacctcg tcttcaagaa gattccttat gaggagttct tcggacaagg
     2941 atggatgaaa ctggaaaaga atgaaaggac cccttatatc atgaaaacca ctaagcactt
     3001 caatgacatc agtaacttga ttgcttcaga aatcatccgc aatgaggaca tcaacgccag
     3061 ggtgagcgcc atcgagaagt gggtggccgt agctgacata tgccgctgcc tccacaacta
     3121 caatgccgta ctggagatca cctcgtccat gaaccgcagt gcaatcttcc ggctcaaaaa
     3181 gacgtggctc aaagtctcta agcagactaa agctttgatt gataagctcc aaaagcttgt
     3241 gtcatctgag ggcagattta agaatctcag agaagctctg aaaaattgtg acccaccctg
     3301 tgtcccttac ctggggatgt acctcaccga cctggccttc atcgaggagg ggacgcccaa
     3361 ttacacggaa gacggcctgg tcaacttctc caagatgagg atgatatccc atattatccg
     3421 agagattcgc cagtttcaac aaactgccta caaaatagag caccaagcaa aggtaacgca
     3481 atatttactg gaccaatctt ttgtaatgga tgaagaaagc ctctacgagt cttctctccg
     3541 aatagaacca aaactcccca cctaggtcaa ctaacttaag ctagcaacgg tttccctcta
     3601 gcgggatcaa ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     3661 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     3721 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     3781 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     3841 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     3901 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     3961 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     4021 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     4081 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     4141 cacggggacg tggttttcct ttgaaaaaca cgataatacc atgaccgagt acaagcccac
     4201 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4261 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4321 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4381 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4441 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4501 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4561 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4621 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4681 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4741 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ctagagttaa catcgaggga
     4801 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4861 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4921 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     4981 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5041 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5101 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5161 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     5221 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5281 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     5341 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     5401 cgcatcgata ccgtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     5461 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     5521 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     5581 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc
     5641 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
     5701 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     5761 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     5821 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     5881 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     5941 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6001 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6061 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
     6121 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6181 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6241 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6301 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     6361 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     6421 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6481 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6541 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6601 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6661 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6721 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6781 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     6841 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6901 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     6961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     7021 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     7081 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     7141 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     7201 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     7261 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
     7321 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
     7381 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     7441 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     7501 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     7561 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     7621 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
     7681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     7801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     7861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     7921 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     7981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     8041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     8101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     8161 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     8221 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     8281 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     8341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     8401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     8461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     8521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     8581 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     8641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     8701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     8761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     8821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8881 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8941 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     9001 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     9061 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     9121 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     9181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     9241 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     9301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     9361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     9421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     9481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     9541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     9601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     9661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     9721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     9781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     9841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    10021 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    10081 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
    10141 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
    10201 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
    10261 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
    10321 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
    10381 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
    10441 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
    10501 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
    10561 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
    10621 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
    10681 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
    10741 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
    10801 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
    10861 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
    10921 gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc
    10981 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    11041 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    11101 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
    11161 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
    11221 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
    11281 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
    11341 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
    11401 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
    11461 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
    11521 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
    11581 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
    11641 gggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
    11701 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
    11761 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
    11821 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
    11881 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
    11941 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
    12001 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
    12061 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
    12121 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
    12181 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
    12241 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
    12301 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
    12361 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
    12421 ccgaggggac ccgacaggcc cgaagga
//
