LOCUS       PB_TetON_mCherry        9019 bp    DNA     circular SYN 05-APR-2025
DEFINITION  For stable clone generation of cells expressing the dCas9-APEX2 
            fusion protein for Genomic locus proteomics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9019)
  AUTHORS   Myers SA, Wright J, Peckner R, Kalish BT, Zhang F, Carr SA
  TITLE     Discovery of proteins associated with a predefined genomic locus via
            dCas9-APEX-mediated proximity labeling.
  JOURNAL   Nat Methods. 2018 May 7. pii: 10.1038/s41592-018-0007-1. doi: 
            10.1038/s41592-018-0007-1.
  PUBMED    29735997
REFERENCE   2  (bases 1 to 9019)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 13, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2018 May 7. pii: 10.1038/s41592-018-0007-1. doi: 
            10.1038/s41592-018-0007-1."
COMMENT     Sequence Label: PB_TetON_mCherry_P2AT2A_GFP-nb-2xKS_FtoG
FEATURES             Location/Qualifiers
     source          1..9019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2384..2434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2435..2485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   147..209
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        658..1036
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    666..684
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    702..720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    738..756
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    774..792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    810..828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    846..864
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    882..900
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     945..969
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1034..1073
                     /label=HN285
     regulatory      join(1049..1054,1055..1057)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1055..1762
                     /codon_start=1
                     /label=mCherry (full)
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1374..1394)
                     /label=backbone_R
     primer_bind     complement(1738..1789)
                     /label=HN423
     misc_feature    1738..1762
                     /label=Feature 26
     misc_feature    1763..1789
                     /label=Feature 28
     misc_feature    1772..1891
                     /label=Feature 31
     primer_bind     1772..1891
                     /label=HN425
     CDS             1772..1828
                     /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             1838..1891
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(1872..1913)
                     /label=HN440
     misc_feature    1872..1891
                     /label=Feature 32
     primer_bind     1874..1943
                     /label=HN424
     misc_feature    1874..1918
                     /label=Feature 30
     primer_bind     1874..1917
                     /label=HN441
     CDS             1892..1918
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1919..1943
                     /label=Feature 29
     CDS             1919..1939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1940..1969
                     /codon_start=1
                     /label=GGGGSx2-linker1
                     /translation="GGGGSGGGGS"
     CDS             1970..2353
                     /codon_start=1
                     /label=GFP_nanobody
                     /translation="MAQVQLVESGGRLVQAGDSLRLSCAASGRTFSTSAMAWFRQAPGR
                     EREFVAAITWTVGNTILGDSVKGRFTISRDRAKNTVDLQMDNLEPEDTAVYYCSARSRG
                     YVLSVLRSVDSYDYWGQGTQVTVS"
     CDS             2354..2383
                     /codon_start=1
                     /label=GGGGSx2-linker2
                     /translation="GGGGSGGGGS"
     CDS             2384..2434
                     /codon_start=1
                     /label=KS
                     /translation="MMMMNKLVLAQGGGSCL"
     misc_feature    2384..2396
                     /label=Feature 35
     primer_bind     complement(2412..2517)
                     /label=HN527
     misc_feature    2412..2434
                     /label=Feature 48
     CDS             2435..2485
                     /codon_start=1
                     /label=KS
                     /translation="MMMMNKLVLAQGGGSCL"
     misc_feature    2477..2485
                     /label=Feature 36
     primer_bind     complement(2536..2559)
                     /label=HN527
     primer_bind     complement(2583..2602)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2627..2675
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        2906..3117
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3130..3398
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3432..4175
                     /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             4176..4229
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(4210..4228)
                     /label=HN440
     primer_bind     4212..4228
                     /label=HN424
     primer_bind     4212..4228
                     /label=HN441
     CDS             4230..4829
                     /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(4230..4249)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4726..4746
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4862..4983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4899..4918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4953..4972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5746..5780
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6271..6287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6271..6287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6284..6306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6295..6311
                     /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(6319..6325,6326..6343,6344..6349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6364..6385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6502..6519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6673..7261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6753..6772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7432..8223,8224..8292))
                     /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     8055..8074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8293..8397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8423..8878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8560..8581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8772..8791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(9005..9019,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(9019,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg
       61 ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca
      121 cgctgcagaa cacgcagcta gattaaccct agaaagataa tcatattgtg acgtacgtta
      181 aagataatca tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt
      241 tattaatttg aatagatatt aagttttatt atatttacac ttacatacta ataataaatt
      301 caacaaacaa tttatttatg tttatttatt tattaaaaaa aacaaaaact caaaatttct
      361 tctataaagt aacaaaactt ttatgaggga cagccccccc ccaaagcccc cagggatgta
      421 attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg
      481 gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt
      541 ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg
      601 ggggatacgg ggaaaaggcc tccacggcca ctagtctttc gtcttcaaga attcctcgag
      661 tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga
      721 acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat
      781 cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag
      841 tttactccct atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga
      901 acgtataagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac
      961 cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     1021 ttcctaccct cgtaaaggtc tagagctagc caccatggtt tccaaaggag aggaggataa
     1081 tatggctata attaaagagt ttatgcggtt caaggtgcat atggagggat ctgtgaacgg
     1141 tcacgagttc gaaatcgagg gcgaaggcga ggggcgaccc tatgaaggga cacagacggc
     1201 caaactgaag gtgaccaagg gtggccccct gcctttcgcc tgggacatcc tgtcacccca
     1261 gtttatgtac ggctctaagg cttacgttaa gcaccctgcc gatattccgg actacttgaa
     1321 actgagcttc cctgaagggt tcaagtggga acgggtaatg aattttgagg atggcggtgt
     1381 agtcacagtt acccaggaca gctctcttca ggacggagaa tttatctata aggttaaact
     1441 tcggggcact aacttcccat cagacggccc cgtgatgcag aagaaaacta tgggctggga
     1501 agccagctca gaacgcatgt atcccgagga cggagcactg aagggagaaa tcaagcagcg
     1561 actcaagctg aaggatgggg gacattatga tgctgaggtc aaaaccacct ataaggccaa
     1621 gaaacccgtt cagcttcctg gggcatacaa cgtgaatatc aaactggata ttacatccca
     1681 taacgaagac tataccatcg tggaacagta cgagcgggcc gagggcagac atagcacagg
     1741 gggcatggat gaattgtaca agggaagcgg agctactaac ttcagcctgc tgaagcaggc
     1801 tggagacgtg gaggagaacc ctggacctgg aagcggagag ggcagaggaa gtcttctaac
     1861 atgcggtgac gtggaggaga atcccggccc atatccgtac gacgtaccag actacgcacc
     1921 taagaagaag aggaaggttg ggggaggcgg cagtggcgga gggggatcta tggctcaggt
     1981 gcagctggtg gaatctggcg gcagactggt gcaggccggc gatagcctga gactgtcttg
     2041 tgccgccagc ggcagaacct tcagcacatc tgccatggcc tggttcagac aggcccctgg
     2101 ccgcgagagg gaatttgtgg ccgccatcac atggaccgtg ggcaacacca tcctgggcga
     2161 cagcgtgaag ggccggttca ccatcagccg ggacagagcc aagaacaccg tggacctcca
     2221 gatggacaac ctggaacccg aggacaccgc cgtgtactac tgctccgcca gatccagagg
     2281 ctacgtgctg tccgtgctgc ggagcgtgga cagctacgat tattggggcc agggcaccca
     2341 agtgaccgtg tctggcggcg gaggaagcgg aggcggagga tctatgatga tgatgaataa
     2401 gttggttcta gcgcagggag gaggatcctg tctaatgatg atgatgaaca agttggttct
     2461 tgctcagggc gggggctctt gcctttagtg tacaagaccg gttgaccggt tagtaatgag
     2521 tttaaacggg ggaggaagac cggttgaccg gttagtaatg agtttaaacg ggggaggcta
     2581 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2641 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2701 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2761 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2821 ggcggcaggc cctgccatag cagatctgcg aaatcggatc cagctgaatc taagtggatc
     2881 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     2941 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     3001 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3061 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     3121 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     3181 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     3241 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     3301 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     3361 gtctttgttt cgttttctgt tctgcgccgt tacagatcta agctgtgacc ggcgcctact
     3421 ctagagccgc catgtctaga ctggacaaga gcaaagtcat aaactctgct ctggaattac
     3481 tcaatggagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     3541 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     3601 tcgagatgct ggacaggcat catacccact cctgccccct ggaaggcgag tcatggcaag
     3661 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     3721 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     3781 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     3841 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     3901 aaagagagac acctaccacc gattctatgc ccccacttct gaaacaagca attgagctgt
     3961 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     4021 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     4081 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     4141 ctcttgacga ttttgacctt gacatgctcc ccggggaggg cagaggaagt cttctaacat
     4201 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg
     4261 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4321 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4381 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4441 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4501 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4561 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4621 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4681 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4741 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4801 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgggt
     4861 aaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4981 ttatcatgtc tggaattgac tcaaatgatg tcaattagtc tatcagaagc tcatctggtc
     5041 tcccttccgg gggacaagac atccctgttt aatatttaaa cagcagtgtt cccaaactgg
     5101 gttcttatat cccttgctct ggtcaaccag gttgcagggt ttcctgtcct cacaggaacg
     5161 aagtccctaa agaaacagtg gcagccaggt ttagccccgg aattgactgg attccttttt
     5221 tagggcccat tggtatggct ttttccccgt atccccccag gtgtctgcag gctcaaagag
     5281 cagcgagaag cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc
     5341 ccgcacgctg ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg
     5401 gctcgctgct gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg
     5461 gggctgtccc tgatatctat aacaagaaaa tatatatata ataagttatc acgtaagtag
     5521 aacatgaaat aacaatataa ttatcgtatg agttaaatct taaaagtcac gtaaaagata
     5581 atcatgcgtc attttgactc acgcggtcgt tatagttcaa aatcagtgac acttaccgca
     5641 ttgacaagca cgcctcacgg gagctccaag cggcgactga gatgtcctaa atgcacagcg
     5701 acggattcgc gctatttaga aagagagagc aatatttcaa gaatgcatgc gtcaatttta
     5761 cgcagactat ctttctaggg ttaatctagc tgcatcagga tcatatcgtc gggtcttttt
     5821 tccggctcag tcatcgccca agctggcgct atctgggcat cggggaggaa gaagcccgtg
     5881 ccttttcccg cgaggttgaa gcggcatgga aagagtttgc cgaggatgac tgctgctgca
     5941 ttgacgttga gcgaaaacgc acgtttacca tgatgattcg ggaaggtgtg gccatgcacg
     6001 cctttaacgg tgaactgttc gttcaggcca cctgggatac cagttcgtcg cggcttttcc
     6061 ggacacagtt ccggatggtc agcccgaagc gcatcagcaa cccgaacaat accggcgaca
     6121 gccggaactg ccgtgccggt gtgcagatta atgacagcgg tgcggcgctg ggatattacg
     6181 tcagcgagga cgggtatcct ggctggatgc cgcagaaatg gacatggata ccccgtgagt
     6241 tacccggcgg gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6301 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6361 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6421 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6481 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6541 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6601 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6661 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6721 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6781 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6841 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6901 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6961 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7021 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7081 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7141 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7201 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8401 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     8461 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     8521 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     8581 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     8641 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     8701 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     8761 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     8821 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     8881 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     8941 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     9001 ttttcccagt cacgacgtt
//
