LOCUS       sequence-455679-       15105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EcTyr aaRS/tRNA system for TAG suppression with EGFP containing 
            three stop codons for triple suppression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15105)
  AUTHORS   Osgood AO, Zheng Y, Roy SJS, Biris N, Hussain M, Loynd C, Jewel D, 
            Italia JS, Chatterjee A
  TITLE     An Efficient Opal-Suppressor Tryptophanyl Pair Creates New Routes 
            for Simultaneously Incorporating up to Three Distinct Noncanonical 
            Amino Acids into Proteins in Mammalian Cells.
  JOURNAL   Angew Chem Int Ed Engl. 2023 May 2;62(19):e202219269. doi: 
            10.1002/anie.202219269. Epub 2023 Apr 12.
  PUBMED    36905325
REFERENCE   2  (bases 1 to 15105)
  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"; journalName: "Angew Chem 
            Int Ed Engl."; date: "2023-05-2"; volume: "62(19):e202219269. doi"; 
            pages: " 10.1002/anie.202219269. Epub 2023 Apr 12"
COMMENT     Sequence Label: 
            pAcBac3-OMeYRS-8xYtR-EGFP-3TAA-40TGA-152TAG-TEV-3xUAA
FEATURES             Location/Qualifiers
     source          1..15105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      168..622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(255..274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     464..485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        648..752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(753..821,822..1613)
                     /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(971..990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1784..2372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2273..2292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2526..2543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2677..2901
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2968..3501)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3690..3695,3696..3712,3713..3718))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4148..4372
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4361..4378
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     regulatory      5723..5732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5751..5770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5752..5770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5823..6222)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     promoter        complement(6332..6426)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    6332..6350
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6360..6378
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(6516..6764)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(6874..6968)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    6874..6892
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6902..6920
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(7058..7306)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(7416..7510)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    7416..7434
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7444..7462
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(7600..7848)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(7958..8052)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    7958..7976
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7986..8004
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(8142..8390)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     enhancer        8554..8857
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          9136..10152
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10076..10098
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10160..10179
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             10229..10939
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT*GKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV*IMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(10268..10289)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10529..10548)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10876..10897
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             10949..10978
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             10994..11017
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             11018..11038
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             11048..11755
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     misc_feature    11815..12403
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11868..11888)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12286..12297)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12471..12490)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    12536..12591
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(12590..12609)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        12943..13034
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     12959..12976
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(13093..13140,13141..14628)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     polyA_signal    14761..14895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14798..14817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14852..14871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  14924..15089
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc
       61 tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt
      121 aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag
      181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
      301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
      361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
      421 gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa
      481 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
      541 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      601 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
      661 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
      721 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
      781 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
      841 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
      901 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
      961 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1021 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1081 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1141 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1201 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1261 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1321 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1381 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1441 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1501 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1561 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1621 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1681 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     1741 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1801 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1861 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1921 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1981 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2041 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2101 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2161 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2221 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2281 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2341 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2401 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2461 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2521 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     2581 cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta
     2641 cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa
     2701 actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag aatagttgta
     2761 aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag
     2821 caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag
     2881 ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg
     2941 gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg
     3001 catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta
     3061 atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt
     3121 gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat
     3181 atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac
     3241 aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc
     3301 gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc
     3361 gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg
     3421 aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac
     3481 atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
     3541 aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag
     3601 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
     3661 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact
     3721 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg
     3781 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc
     3841 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct
     3901 tgcttcagga gatcggtaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg
     3961 atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact
     4021 ctagctatag ttctagtggt tggctacgta cccgtagtgg ctatggcagg gcttgcgctt
     4081 aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc
     4141 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     4201 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     4261 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     4321 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     4381 gatcagcggg tttaaacggg ccctctagac tcgagttaaa gtcgacttaa cgcgttgaat
     4441 tcttaaacgg gccctttcca gcaaatcaga cagtaattct ttttaccgcg acgcagtaag
     4501 gtaaaacgac caaacagacg atcttcttct ttaaagaagt attcaggatc ggactgtttt
     4561 tcaccgttaa tggtgatggc attggaggcg atagttttac gtgcctgacc acgggaaggt
     4621 tgcagttcag aatcgaccag tgcctgcatc aggtctgcgc ccttttccat ctcaaccatc
     4681 ggtacgccgt cctgcgccag ctgttcgaag tccgcttcac tcagcgcact caaagaaccg
     4741 ctgaacaggc attcggtaat acgttttgcc gcctgtaaac cttcttcacc gtgaaccaga
     4801 cgagtcacct gctccgccag tacatactgg gcgcgcggtg ctttaccgct gtttttatct
     4861 tcttcttcca gggcgttgat ctcttcaatg ctcataaagg tgaagaactt caggaagcgg
     4921 taaacgtcgg cacgcgcagt gttgatccag aactggtaga atttgtacgg gctggttttc
     4981 ttcggatcca accagactgc gccgccttca gttttaccaa atttggtgcc atctgcttta
     5041 gtgatcagcg gaacggtcag gccaaacacc tgattctgat gcagacgacg ggtcaggtcg
     5101 ataccagaag tgatgttacc ccactggtca gaaccaccaa tttgcagcac cacaccgtac
     5161 tgtttgttca aacaggccat actataaccc tgcagcaggt tgtaggaaaa ctcagtgaac
     5221 gaaatcccct gatcttcacg gttgagacgc tgcttaaccg cttctttgtt gatcatctgg
     5281 ttaacggaga agtgtttgcc aatatcgcgc aggaaggtca gcacattcat attgccgaac
     5341 cagtcataat tattggccgc gatagcagag ttttctccac agtcgaaatc gaggaacggg
     5401 gcaacctgct tacggatttt gtccacccac tcctgaacag tttcttcggt gttcagctta
     5461 cgctcggcag ctttgaagct cgggtcgcca atcagacccg tcgcgccgcc taccagcgca
     5521 accggcttgt ggcccgcctg ctggaagcgt ttcaggcata acaatggaac aagatgcccc
     5581 aaatgcaagc tgtcagcggt aggatcgaag ccacacacga gtgcgatcgg gccttgcgcc
     5641 agtcgctctg ctaacgcttc ctcgtccgtc acctgggcta ccagcccccg ctcttgcaat
     5701 tgtttaatca agttactgct tgccatggtg gcgctagcca gcttgggtct ccctatagtg
     5761 agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct
     5821 agaccaagtg acgatcacag cgatccacaa acaagaaccg cgacccaaat cccggctgcg
     5881 acggaactag ctgtgccaca cccggcgcgt ccttatataa tcatcggcgt tcaccgcccc
     5941 acggagatcc ctccgcagaa tcgccgagaa gggactactt ttcctcgcct gttccgctct
     6001 ctggaaagaa aaccagtgcc ctagagtcac ccaagtcccg tcctaaaatg tccttctgct
     6061 gatactgggg ttctaaggcc gagtcttatg agcagcgggc cgctgtcctg agcgtccggg
     6121 cggaaggatc aggacgctcg ctgcgccctt cgtctgacgt ggcagcgctc gccgtgagga
     6181 ggggggcgcc cgcgggaggc gccaaaaccc ggcgcggagg ccttcgaacg gccactagca
     6241 aaaaatggag ggggacggat tcgaaccgcc gaacccaaag ggagcggatt tagagtccgc
     6301 cgcgtttagc cacttcgcta cccctccggt gtctctatca ctgataggga acttataagt
     6361 ctctatcact gatagggatt tcacgtttat ggtgatttcc cagaacacat agcgacatgc
     6421 aaatattaaa aaatggtggg ggaaggattc gaaccttcga agtctgtgac ggcagattta
     6481 gagtctgctc cctttggccg ctcgggaacc ccaccggtgt ttcgtccttt ccacaagata
     6541 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
     6601 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
     6661 tgtactaata tctttgtgtt tacagtcaaa ttaattctaa ttatctctct aacagccttg
     6721 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcttcctg cccgacctag
     6781 caaaaaatgg agggggacgg attcgaaccg ccgaacccaa agggagcgga tttagagtcc
     6841 gccgcgttta gccacttcgc tacccctccg gtgtctctat cactgatagg gaacttataa
     6901 gtctctatca ctgataggga tttcacgttt atggtgattt cccagaacac atagcgacat
     6961 gcaaatatta aaaaatggtg ggggaaggat tcgaaccttc gaagtctgtg acggcagatt
     7021 tagagtctgc tccctttggc cgctcgggaa ccccaccggt gtttcgtcct ttccacaaga
     7081 tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt
     7141 taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat
     7201 tttgtactaa tatctttgtg tttacagtca aattaattct aattatctct ctaacagcct
     7261 tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcttcc tgcccgacct
     7321 agcaaaaaat ggagggggac ggattcgaac cgccgaaccc aaagggagcg gatttagagt
     7381 ccgccgcgtt tagccacttc gctacccctc cggtgtctct atcactgata gggaacttat
     7441 aagtctctat cactgatagg gatttcacgt ttatggtgat ttcccagaac acatagcgac
     7501 atgcaaatat taaaaaatgg tgggggaagg attcgaacct tcgaagtctg tgacggcaga
     7561 tttagagtct gctccctttg gccgctcggg aaccccaccg gtgtttcgtc ctttccacaa
     7621 gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat
     7681 tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt
     7741 attttgtact aatatctttg tgtttacagt caaattaatt ctaattatct ctctaacagc
     7801 cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctctt cctgcccgac
     7861 ctagcaaaaa atggaggggg acggattcga accgccgaac ccaaagggag cggatttaga
     7921 gtccgccgcg tttagccact tcgctacccc tccggtgtct ctatcactga tagggaactt
     7981 ataagtctct atcactgata gggatttcac gtttatggtg atttcccaga acacatagcg
     8041 acatgcaaat attaaaaaat ggtgggggaa ggattcgaac cttcgaagtc tgtgacggca
     8101 gatttagagt ctgctccctt tggccgctcg ggaaccccac cggtgtttcg tcctttccac
     8161 aagatatata aagccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc
     8221 attttaaaac ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac
     8281 gtattttgta ctaatatctt tgtgtttaca gtcaaattaa ttctaattat ctctctaaca
     8341 gccttgtatc gtatatgcaa atatgaagga atcatgggaa ataggccctc ttcctgcccg
     8401 acctagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa
     8461 aacggatcct gcaggctgat cactgcttga gcctagttat taatagtaat caattacggg
     8521 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     8581 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     8641 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     8701 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     8761 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     8821 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
     8881 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
     8941 attattttgt gcagcgatgg gggcgggggg gggggggggc gcgcgccrgg sggggsgggg
     9001 sggggsgrgg gggggggggg ggsgrggsgg rgaggkgcgg cggcagccaa tcagagcggc
     9061 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
     9121 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
     9181 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
     9241 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
     9301 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
     9361 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
     9421 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
     9481 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc
     9541 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg
     9601 caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     9661 ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     9721 gtgccgggcg gggcggggcc gcctcgggcc ggggrgggct cgggggaggg gcgcggcggc
     9781 ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa
     9841 tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg
     9901 aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag
     9961 gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag
    10021 cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
    10081 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
    10141 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
    10201 gaattggcca aggaggccca tggtgtagag caagggcgag gagctgttca ccggggtggt
    10261 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
    10321 gggcgagggc gatgccactt gaggcaagct gaccctgaag ttcatctgca ccaccggcaa
    10381 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
    10441 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
    10501 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
    10561 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
    10621 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctaaat
    10681 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
    10741 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
    10801 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
    10861 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
    10921 catggacgag ctgtacaagg ggcccttcga acaaaaactc atctcagaag aggatctgaa
    10981 tatgcatacc ggtcatcatc accatcacca tcatcacgaa aatctttatt ttcaaggtgg
    11041 aggaagtgga gaaagtttgt ttaaggggcc gcgtgattac aacccgatat cgagcaccat
    11101 ttgtcatttg acgaatgaat ctgatgggca cacaacatcg ttgtatggta ttggatttgg
    11161 tcccttcatc attacaaaca agcacttgtt tagaagaaat aatggaacac tgttggtcca
    11221 atcactacat ggtgtattca aggtcaagaa caccacgact ttgcaacaac acctcattga
    11281 tgggagggac atgataatta ttcgcatgcc taaggatttc ccaccatttc ctcaaaagct
    11341 gaaatttaga gagccacaaa gggaagagcg catatgtctt gtgacaacca acttccaaac
    11401 taagagcatg tctagcatgg tgtcagacac tagttgcaca ttcccttcat ctgatggcat
    11461 attctggaag cattggattc aaaccaagga tgggcagtgt ggcagtccat tagtatcaac
    11521 tagagatggg ttcattgttg gtatacactc agcatcgaat ttcaccaaca caaacaatta
    11581 tttcacaagc gtgccgaaaa acttcatgga attgttgaca aatcaggagg cgcagcagtg
    11641 ggttagtggt tggcgattaa atgctgactc agtattgtgg gggggccata aagttttcat
    11701 ggtgaaacct gaagagcctt ttcagccagt taaggaagcg actcaactca tgaatcgtcg
    11761 tcgccgtcgc taataataag gcctctaagg ccgaattcaa cgcgttaagt cgacaatcaa
    11821 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
    11881 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
    11941 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
    12001 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
    12061 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
    12121 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
    12181 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
    12241 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
    12301 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
    12361 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaactcggc
    12421 cagcacagtg gtcgatcgac caatgccctg gctcacaaat accactgaga tctttttccc
    12481 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
    12541 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
    12601 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
    12661 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
    12721 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
    12781 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
    12841 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
    12901 cttctttgcg gccgcggtcc gtatactccg gaatattaat agatcatgga gataattaaa
    12961 atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa
    13021 aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc gaactcctaa
    13081 aaaaccgcca ccatgaagtg ccttttgtac ttagcctttt tattcattgg ggtgaattgc
    13141 aagttcacca tagtttttcc acacaaccaa aaaggaaact ggaaaaatgt tccttctaat
    13201 taccattatt gcccgtcaag ctcagattta aattggcata atgacttaat aggcacagcc
    13261 ttacaagtca aaatgcccaa gagtcacaag gctattcaag cagacggttg gatgtgtcat
    13321 gcttccaaat gggtcactac ttgtgatttc cgctggtatg gaccgaagta tataacacat
    13381 tccatccgat ccttcactcc atctgtagaa caatgcaagg aaagcattga acaaacgaaa
    13441 caaggaactt ggctgaatcc aggcttccct cctcaaagtt gtggatatgc aactgtgacg
    13501 gatgccgaag cagtgattgt ccaggtgact cctcaccatg tgctggttga tgaatacaca
    13561 ggagaatggg ttgattcaca gttcatcaac ggaaaatgca gcaattacat atgccccact
    13621 gtccataact ctacaacctg gcattctgac tataaggtca aagggctatg tgattctaac
    13681 ctcatttcca tggacatcac cttcttctca gaggacggag agctatcatc cctgggaaag
    13741 gagggcacag ggttcagaag taactacttt gcttatgaaa ctggaggcaa ggcctgcaaa
    13801 atgcaatact gcaagcattg gggagtcaga ctcccatcag gtgtctggtt cgagatggct
    13861 gataaggatc tctttgctgc agccagattc cctgaatgcc cagaagggtc aagtatctct
    13921 gctccatctc agacctcagt ggatgtaagt ctaattcagg acgttgagag gatcttggat
    13981 tattccctct gccaagaaac ctggagcaaa atcagagcgg gtcttccaat ctctccagtg
    14041 gatctcagct atcttgctcc taaaaaccca ggaaccggtc ctgctttcac cataatcaat
    14101 ggtaccctaa aatactttga gaccagatac atcagagtcg atattgctgc tccaatcctc
    14161 tcaagaatgg tcggaatgat cagtggaact accacagaaa gggaactgtg ggatgactgg
    14221 gcaccatatg aagacgtgga aattggaccc aatggagttc tgaggaccag ttcaggatat
    14281 aagtttcctt tatacatgat tggacatggt atgttggact ccgatcttca tcttagctca
    14341 aaggctcagg tgttcgaaca tcctcacatt caagacgctg cttcgcaact tcctgatgat
    14401 gagagtttat tttttggtga tactgggcta tccaaaaatc caatcgagct tgtagaaggt
    14461 tggttcagta gttggaaaag ctctattgcc tcttttttct ttatcatagg gttaatcatt
    14521 ggactattct tggttctccg agttggtatc catctttgca ttaaattaaa gcacaccaag
    14581 aaaagacaga tttatacaga catagagatg aaccgacttg gaaagtgata aaagcttgtc
    14641 gagaagtact agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
    14701 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
    14761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    14821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    14881 tatcatgtct ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaat
    14941 ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag
    15001 ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc
    15061 ccagttccca actattttgt ccgcccacag cggggcattt ttctt
//
