LOCUS       sequence-387472-        9370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-On cilia-targeted RhoA sensor (Xlone piggybac back-bone); 
            HTR6-fused with a sGFP-mScarlet-I based sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9370)
  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 9370)
  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-HTR6-RhoA sensor
FEATURES             Location/Qualifiers
     source          1..9370
                     /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"
     regulatory      389..398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2024..2719
                     /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(2157..2175)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2425..2444
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(2915..2917,2918..3430)
                     /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(2957..2978)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3218..3237)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3565..3586
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3605..4183
                     /codon_start=1
                     /product="human Rho family small GTPase RhoA"
                     /label=RhoA
                     /translation="AAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADI
                     EVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKH
                     FCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTK
                     DGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    4367..4501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4777..4811
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4822..4839)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4822..4838)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4831..4853)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5047..5066)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5166..5188
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5226..5244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5312..5416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5417..5485,5486..6277)
                     /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(5635..5654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6448..7036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6937..6956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7190..7207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7324..7345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7360..7365,7366..7383,7384..7390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7398..7414
                     /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     7403..7425
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7422..7438
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7422..7438
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   7464..7526
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     gap             7589..7631
                     /estimated_length=43
     polyA_signal    7667..7748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7865..8263)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(8264..8320)
                     /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(8330..9073)
                     /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(9074..9097)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      9097..9106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9126..9337)
                     /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 caccatggtc ccagagccgg gcccaaccgc
      421 caatagcacc ccggcctggg gggcagggcc gccgtcggcc ccggggggca gcggctgggt
      481 ggcggccgcg ctgtgcgtgg tcatcgcgct gacggcggcg gccaactcgc tgctgatcgc
      541 gctcatctgc actcagcccg cgctgcgcaa cacgtccaac ttcttcctgg tgtcgctctt
      601 cacgtctgac ctgatggtgg ggctggtggt gatgccgccg gccatgctga acgcgctgta
      661 cgggcgctgg gtgctggcgc gcggcctctg cctgctctgg accgccttcg acgtgatgtg
      721 ctgcagcgcc tccatcctca acctctgcct catcagcctg gaccgctacc tgctcatcct
      781 ctcgccgctg cgctacaagc tgcgcatgac gcccctgcgt gccctggccc tagtcctggg
      841 cgcctggagc ctcgccgctc tcgcctcctt cctgcccctg ctgctgggct ggcacgagct
      901 gggccacgca cggccacccg tccctggcca gtgccgcctg ctggccagcc tgccttttgt
      961 ccttgtggcg tcgggcctca ccttcttcct gccctcgggt gccatatgct tcacctactg
     1021 caggatcctg ctagctgccc gcaagcaggc cgtgcaggtg gcctccctca ccaccggcat
     1081 ggccagtcag gcctcggaga cgctgcaggt gcccaggacc ccacgcccag gggtggagtc
     1141 tgctgacagc aggcgtctag ccacgaagca cagcaggaag gccctgaagg ccagcctgac
     1201 gctgggcatc ctgctgggca tgttctttgt gacctggttg cccttctttg tggccaacat
     1261 agtccaggcc gtgtgcgact gcatctcccc aggcctcttc gatgtcctca catggctggg
     1321 ttactgtaac agcaccatga accccatcat ctacccactc ttcatgcggg acttcaagcg
     1381 ggcgctgggc aggttcctgc catgtccacg ctgtccccgg gagcgccagg ccagcctggc
     1441 ctcgccatca ctgcgcacct ctcacagcgg cccccggccc ggccttagcc tacagcaggt
     1501 gctgccgctg cccctgccgc cggactcaga ttcggactca gacgcaggct caggcggctc
     1561 ctcgggcctg cggctcacgg cccagctgct gcttcctggc gaggccaccc aggacccccc
     1621 gctgcccacc agggccgctg ccgccgtcaa tttcttcaac atcgaccccg cggagcccga
     1681 gctgcggccg catccacttg gcatccccac gaacaacggt acagcgggcc cgggatccgc
     1741 gatcgccacc agtctgggcc cagtcgagct tctgttgcga gggagcagca ggagattgga
     1801 tctgcttcac caacagttgc aagaattgca cgctcatgga agcggctctc tccttgagca
     1861 attgggtctt gcgggagcgg atttggcggc tcccggcgtg cagcaacaat tggaattgga
     1921 gcgggagcga ctgcggcggg agatacggaa agaactcaaa ttgaaggagg gcgcagagaa
     1981 ccttagacgc gccaccactg atcttggacg gggttcagga tccatggtga gcaagggcga
     2041 ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca tgaacggcca
     2101 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2161 gctgaaggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt cccctcagtt
     2221 catgtacggc tccagggcct tcatcaagca ccccgccgac atccccgact actataagca
     2281 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt
     2341 gaccgtgacc caggacacct ccctggagga cggcaccctg atctacaagg tgaagctccg
     2401 cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg gctgggaagc
     2461 gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta agatggccct
     2521 gcgcctgaag gacggcggcc gctacctggc ggacttcaag accacctaca aggccaagaa
     2581 gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag ttggacatca cctcccacaa
     2641 cgaggactac accgtggtgg aacagtacga acgctccgag ggccgccact ccaccggcgg
     2701 catggacgag ctgtacaagt ccggactcag atctaacatc gaggaagcac aaaagcagaa
     2761 ggaacagaga caggcagcag aggagctggc aaacggatct gaagcacaaa agcagaagga
     2821 acagagacag gcagcagagg agctggcaaa cggatctgaa gcacaaaagc agaaggaaca
     2881 gagacaggca gcagaggagc tggcaaacgg atccgtgagc aagggcgagg agctgttcac
     2941 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3001 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3061 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3121 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3181 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3241 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3301 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3361 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     3421 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3481 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     3541 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccggggc
     3601 tagcgctgcc atccggaaga aactggtgat tgttggtgat ggagcctgtg gaaagacatg
     3661 cttgctcata gtcttcagca aggaccagtt cccagaggtg tatgtgccca cagtgtttga
     3721 gaactatgtg gcagatatcg aggtggatgg aaagcaggta gagttggctt tgtgggacac
     3781 agctgggcag gaagattatg atcgcctgag gcccctctcc tacccagata ccgatgttat
     3841 actgatgtgt ttttccatcg acagccctga tagtttagaa aacatcccag aaaagtggac
     3901 cccagaagtc aagcatttct gtcccaacgt gcccatcatc ctggttggga ataagaagga
     3961 tcttcggaat gatgagcaca caaggcggga gcttgccaag atgaagcagg agccggtgaa
     4021 acctgaagaa ggcagagata tggcaaacag gattggcgct tttgggtaca tggagtgttc
     4081 agcaaagacc aaagatggag tgagagaggt ttttgaaatg gctacgagag ctgctctgca
     4141 agctagacgt gggaagaaaa aatctgggtg ccttgtcttg tgataaacta gtagaccacc
     4201 tcccctgcga gctaagctgg acagccaatg acgggtaaga gagtgacatt tttcactaac
     4261 ctaagacagg agggccgtca gagctactgc ctaatccaaa gacgggtaaa agtgataaaa
     4321 atgtatcact ccaacctaag acaggcgcag cttccgaggg atttgagatc cagacatgat
     4381 aagatacatt gatgagtttg gacaaaccaa aactagaatg cagtgaaaaa aatgccttat
     4441 ttgtgaaatt tgtgatgcta ttgccttatt tgtaaccatt ataagctgca ataaacaagt
     4501 ttgatatcta taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa
     4561 taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt
     4621 cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc
     4681 acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg
     4741 cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta
     4801 tctttctagg gttaagaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4861 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4921 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4981 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     5041 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5101 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5161 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5221 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5281 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5341 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5401 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5461 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5521 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5581 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5641 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5701 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5761 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5821 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5881 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5941 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6001 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6061 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6121 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6181 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6241 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6301 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6361 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6421 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6481 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6541 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6601 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6661 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6721 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6781 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6841 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6901 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6961 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7021 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7081 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7141 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     7201 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     7261 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     7321 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     7381 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     7441 tgattacgcc aaggtcgact taaccctaga aagataatca tattgtgacg tacgttaaag
     7501 ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat
     7561 taatttgaat agatattaag ttttattann nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     7621 nnnnnnnnnn nmtmaaaakw tcttctataa agtaacaaaa cttttagcag tgaaaaaaat
     7681 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     7741 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     7801 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctgg
     7861 agatttagcc ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg
     7921 gctgtccatc actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct
     7981 gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac
     8041 aagtcaggtt gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag
     8101 gtcccccagt aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag
     8161 ctgcgctggc gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc
     8221 tttcaatgag ggtggattct tcttgagaca aaggcttggc catggggccg gggttctcct
     8281 ccacgtcgcc ggcctgcttc agcaggctga agttggtggc gccgctgccc ccggggagca
     8341 tgtcaaggtc aaaatcgtca agagcgtcag caggcagcat atcaaggtca aagtcgtcaa
     8401 gggcatcggc tgggagcatg tctaagtcaa aatcgtcaag ggcgtcggtc ggcccgccgc
     8461 tttcgcactt tagctgtttc tccaggccac atatgattag ttccaggccg aaaaggaagg
     8521 caggttcggc tccctgccgg tcgaacagct caattgcttg tttcagaagt gggggcatag
     8581 aatcggtggt aggtgtctct ctttcctctt ttgctacttg atgctcctgt tcctccaata
     8641 cgcagcccag tgtaaagtgg cccacggcgg acagagcgta cagtgcgttc tccagggaga
     8701 agccttgctg acacaggaac gcgagctgat tttccagggt ttcgtactgt ttctctgttg
     8761 ggcgggtgcc gagatgcact ttagccccgt cgcgatgtga gaggagagca cagcggtatg
     8821 acttggcgtt gttccgcaga aagtcttgcc atgactcgcc ttccaggggg caggagtggg
     8881 tatgatgcct gtccagcatc tcgattggca gggcatcgag cagggcccgc ttgttcttca
     8941 cgtgccagta cagggtaggc tgctcaactc ccagcttttg agcgagtttc cttgtcgtca
     9001 ggccttcgat accgactcca ttgagtaatt ccagagcaga gtttatgact ttgctcttgt
     9061 ccagtctaga catcttatcg tcatcgtctt tgtaatccat ggtggcggat cccgcgtcac
     9121 gacacctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca
     9181 cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt
     9241 tcccagttta ccccgcgcca ccttctctag gcaccggttc aattgccgac ccctcccccc
     9301 aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagccactc
     9361 gagtggaatt
//
