LOCUS       sequence-387515-        9369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-On cilia-targeted RhoA sensor (Xlone piggybac back-bone); 
            Arl13b-fused with a sGFP-mScarlet-I based sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9369)
  AUTHORS   Sheu SH, Upadhyayula S, Dupuy V, Pang S, Deng F, Wan J, Walpita D, 
            Pasolli HA, Houser J, Sanchez-Martinez S, Brauchi SE, Banala S, 
            Freeman M, Xu CS, Kirchhausen T, Hess HF, Lavis L, Li Y, 
            Chaumont-Dubel S, Clapham DE
  TITLE     A serotonergic axon-cilium synapse drives nuclear signaling to alter
            chromatin accessibility.
  JOURNAL   Cell. 2022 Sep 1;185(18):3390-3407.e18. doi: 
            10.1016/j.cell.2022.07.026.
  PUBMED    36055200
REFERENCE   2  (bases 1 to 9369)
  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.cell.2022.07"; journalName: "Cell"; date: "2022-09-1- 1";
            volume: "185"; issue: "18"; pages: "3390-3407.e18"
COMMENT     Sequence Label: pB-Tet-On-Arl13b-RhoA sensor
FEATURES             Location/Qualifiers
     source          1..9369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..376
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    53..71
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    89..107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1988..2683
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2121..2139)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2389..2408
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(2879..2881,2882..3394)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(2921..2942)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3182..3201)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3529..3550
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3569..4147
                     /codon_start=1
                     /product="human Rho family small GTPase RhoA"
                     /label=RhoA
                     /translation="AAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADI
                     EVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKH
                     FCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTK
                     DGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    4331..4465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4741..4775
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4786..4803)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4786..4802)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4795..4817)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5011..5030)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5130..5152
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5190..5208)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5276..5380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5381..5449,5450..6241)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5599..5618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6412..7000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6901..6920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7154..7171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7288..7309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7324..7329,7330..7347,7348..7354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7362..7378
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7367..7389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7386..7402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7386..7402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   7428..7490
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    7666..7747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7864..8262)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(8263..8319)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(8329..9072)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(9073..9096)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      9096..9105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9125..9336)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
       61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
      121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
      181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      241 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
      301 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      361 ttcctaccct cgtaaaggta ccacgcgtgc caccatgttc agtctgatgg ccagttgctg
      421 cggctggttc aagcggtggc gggagcctgt cagaaaggtg actcttttga tggtgggact
      481 tgataatgct ggtaaaaccg caacagcaaa gggaatccaa ggagaatacc ctgaagatgt
      541 agctcctact gttggatttt caaaaattaa ccttagacaa ggaaagtttg aagtcaccat
      601 ctttgacttg ggaggtggaa taagaattcg gggaatctgg aagaattact atgctgaatc
      661 ctatggggta atatttgttg tggattccag tgatgaagag agaatggaag agacaaaaga
      721 ggctatgtca gaaatgctaa gacatcctag gatatcggga aagcctatat tggtgttggc
      781 aaataaacaa gataaagaag gagctttagg agaagctgat gtcattgaat gtctatctct
      841 ggaaaaattg gtcaatgagc acaagtgcct gtgtcagata gaaccatgtt cagcaatctc
      901 ggggtatgga aagaaaattg acaagtccat taaaaaaggc ctttattggc tgctacatgt
      961 tattgcaaga gactttgatg ccttaaatga acgcatccaa aaagagacaa cagagcagcg
     1021 tgctcttgag gaacaagaga aacaagaaag agctgaacga gtgcgaaaat tacgagaaga
     1081 aagaaaacaa aatgaacagg agcaggctga actcgatgga accagtggtc tggctgagtt
     1141 ggacccagaa ccaacgaatc ctttccagcc aatagcatct gtaatcattg agaatgaagg
     1201 aaaacttgaa agagagaaaa aaaaccaaaa aatggagaaa gacagtgatg gctgccacct
     1261 gaaacataaa atggagcatg agcaaataga gacacaaggc caggttaatc acaatggcca
     1321 aaaaaataat gaatttggac tagtagaaaa ttataaggag gcattaacac agcagttaaa
     1381 gaatgaagat gagacagacc ggccatcatt ggaatcagct aatggtaaaa agaaaactaa
     1441 gaaactaaga atgaaaagga accaccgggt agaaccactt aatatagatg actgtgctcc
     1501 tgagagtcca acgccacccc caccccctcc tcctgttggc tggggaaccc ctaaagtcac
     1561 tagacttcca aaacttgagc ctcttggtga aacacatcat aatgatttct ataggaagcc
     1621 actgcctccc ctggctgtgc cacagcgacc taacagtgat gctcatgatg tgatctcaaa
     1681 cggtacagcg ggcccgggat ccgcgatcgc caccagtctg ggcccagtcg agcttctgtt
     1741 gcgagggagc agcaggagat tggatctgct tcaccaacag ttgcaagaat tgcacgctca
     1801 tggaagcggc tctctccttg agcaattggg tcttgcggga gcggatttgg cggctcccgg
     1861 cgtgcagcaa caattggaat tggagcggga gcgactgcgg cgggagatac ggaaagaact
     1921 caaattgaag gagggcgcag agaaccttag acgcgccacc actgatcttg gacggggttc
     1981 aggatccatg gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca
     2041 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2101 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc
     2161 ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcatca agcaccccgc
     2221 cgacatcccc gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2281 gaacttcgag gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac
     2341 cctgatctac aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca
     2401 gaagaagaca atgggctggg aagcgtccac cgagcggttg taccccgagg acggcgtgct
     2461 gaagggcgac attaagatgg ccctgcgcct gaaggacggc ggccgctacc tggcggactt
     2521 caagaccacc tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg
     2581 caagttggac atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc
     2641 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctaa
     2701 catcgaggaa gcacaaaagc agaaggaaca gagacaggca gcagaggagc tggcaaacgg
     2761 atctgaagca caaaagcaga aggaacagag acaggcagca gaggagctgg caaacggatc
     2821 tgaagcacaa aagcagaagg aacagagaca ggcagcagag gagctggcaa acggatccgt
     2881 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2941 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3001 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3061 gaccaccctg acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     3121 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3181 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3241 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3301 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     3361 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     3421 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3481 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3541 ggagttcgtg accgccgccg gggctagcgc tgccatccgg aagaaactgg tgattgttgg
     3601 tgatggagcc tgtggaaaga catgcttgct catagtcttc agcaaggacc agttcccaga
     3661 ggtgtatgtg cccacagtgt ttgagaacta tgtggcagat atcgaggtgg atggaaagca
     3721 ggtagagttg gctttgtggg acacagctgg gcaggaagat tatgatcgcc tgaggcccct
     3781 ctcctaccca gataccgatg ttatactgat gtgtttttcc atcgacagcc ctgatagttt
     3841 agaaaacatc ccagaaaagt ggaccccaga agtcaagcat ttctgtccca acgtgcccat
     3901 catcctggtt gggaataaga aggatcttcg gaatgatgag cacacaaggc gggagcttgc
     3961 caagatgaag caggagccgg tgaaacctga agaaggcaga gatatggcaa acaggattgg
     4021 cgcttttggg tacatggagt gttcagcaaa gaccaaagat ggagtgagag aggtttttga
     4081 aatggctacg agagctgctc tgcaagctag acgtgggaag aaaaaatctg ggtgccttgt
     4141 cttgtgataa actagtagac cacctcccct gcgagctaag ctggacagcc aatgacgggt
     4201 aagagagtga catttttcac taacctaaga caggagggcc gtcagagcta ctgcctaatc
     4261 caaagacggg taaaagtgat aaaaatgtat cactccaacc taagacaggc gcagcttccg
     4321 agggatttga gatccagaca tgataagata cattgatgag tttggacaaa ccaaaactag
     4381 aatgcagtga aaaaaatgcc ttatttgtga aatttgtgat gctattgcct tatttgtaac
     4441 cattataagc tgcaataaac aagtttgata tctataacaa gaaaatatat atataataag
     4501 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa
     4561 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca
     4621 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt
     4681 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg
     4741 catgcgtcaa ttttacgcag actatctttc tagggttaag aattcactgg ccgtcgtttt
     4801 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4861 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4921 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4981 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5041 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5101 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5161 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5221 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5281 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5341 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5401 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5461 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5521 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5581 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5641 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5701 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5761 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5821 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5881 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5941 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6001 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6061 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6121 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6181 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6241 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6301 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6361 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6421 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6481 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6541 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6601 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6661 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6721 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6781 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6841 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6901 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6961 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7021 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7081 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7141 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7201 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7261 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7321 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7381 tcacacagga aacagctatg accatgatta cgccaaggtc gacttaaccc tagaaagata
     7441 atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg
     7501 gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca
     7561 ctttcttact aataataact tcaacaaaca atttatttat gtttatttat ttattaaaaa
     7621 aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttagcagt gaaaaaaatg
     7681 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     7741 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     7801 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctgga
     7861 gatttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     7921 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     7981 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     8041 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     8101 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     8161 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     8221 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggggccgg ggttctcctc
     8281 cacgtcgccg gcctgcttca gcaggctgaa gttggtggcg ccgctgcccc cggggagcat
     8341 gtcaaggtca aaatcgtcaa gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag
     8401 ggcatcggct gggagcatgt ctaagtcaaa atcgtcaagg gcgtcggtcg gcccgccgct
     8461 ttcgcacttt agctgtttct ccaggccaca tatgattagt tccaggccga aaaggaaggc
     8521 aggttcggct ccctgccggt cgaacagctc aattgcttgt ttcagaagtg ggggcataga
     8581 atcggtggta ggtgtctctc tttcctcttt tgctacttga tgctcctgtt cctccaatac
     8641 gcagcccagt gtaaagtggc ccacggcgga cagagcgtac agtgcgttct ccagggagaa
     8701 gccttgctga cacaggaacg cgagctgatt ttccagggtt tcgtactgtt tctctgttgg
     8761 gcgggtgccg agatgcactt tagccccgtc gcgatgtgag aggagagcac agcggtatga
     8821 cttggcgttg ttccgcagaa agtcttgcca tgactcgcct tccagggggc aggagtgggt
     8881 atgatgcctg tccagcatct cgattggcag ggcatcgagc agggcccgct tgttcttcac
     8941 gtgccagtac agggtaggct gctcaactcc cagcttttga gcgagtttcc ttgtcgtcag
     9001 gccttcgata ccgactccat tgagtaattc cagagcagag tttatgactt tgctcttgtc
     9061 cagtctagac atcttatcgt catcgtcttt gtaatccatg gtggcggatc ccgcgtcacg
     9121 acacctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac
     9181 ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt
     9241 cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca
     9301 acttctcggg gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagccactcg
     9361 agtggaatt
//
