LOCUS       sequence-455604-       15070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EcTyr aaRS/tRNA system for pAAF incorporation 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 15070)
  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 15070)
  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-pAAFRS-8xYtR-EGFP-3TAA-
            40TGA-152TAG-T...
FEATURES             Location/Qualifiers
     source          1..15070
                     /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      5687..5696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5715..5734)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5716..5734)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5787..6186)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     promoter        complement(6296..6390)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    6296..6314
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6324..6342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(6480..6728)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(6838..6932)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    6838..6856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6866..6884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(7022..7270)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(7380..7474)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    7380..7398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7408..7426
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(7564..7812)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(7922..8016)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    7922..7940
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7950..7968
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(8106..8354)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     enhancer        8518..8821
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          9101..10117
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10041..10063
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10125..10144
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             10194..10904
                     /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(10233..10254)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10494..10513)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10841..10862
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             10914..10943
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             10959..10982
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             10983..11003
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             11013..11720
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     misc_feature    11780..12368
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11833..11853)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12251..12262)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12436..12455)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    12501..12556
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(12555..12574)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        12908..12999
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     12924..12941
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(13058..13105,13106..14593)
                     /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    14726..14860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14763..14782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14817..14836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  14889..15054
                     /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 tcgagttatt tccagcaaat cagacagtaa
     4441 ttctttttac cgcgacgcag taaggtaaaa cgaccaaaca gacgatcttc ttctttaaag
     4501 aagtattcag gatcggactg tttttcaccg ttaatggtga tggcattgga ggcgatagtt
     4561 ttacgtgcct gaccacggga aggttgcagt tcagaatcga ccagtgcctg catcaggtct
     4621 gcgccctttt ccatctcaac catcggtacg ccgtcctgcg ccagctgttc gaagtccgct
     4681 tcactcagcg cactcaaaga accgctgaac aggcattcgg taatacgttt tgccgcctgt
     4741 aaaccttctt caccgtgaac cagacgagtc acctgctccg ccagtacata ctgggcgcgc
     4801 ggtgctttac cgctgttttt atcttcttct tccagggcgt tgatctcttc aatgctcata
     4861 aaggtgaaga acttcaggaa gcggtaaacg tcggcacgcg cagtgttgat ccagaactgg
     4921 tagaatttgt acgggctggt tttcttcgga tctaaccaga ctgcgccgcc ttcagtttta
     4981 ccaaatttgg tgccatctgc tttagtgatc agcggaacgg tcaggccaaa cacctgattc
     5041 tgatgcagac gacgggtcag gtcgatacca gaagtgatgt taccccactg gtcagaacca
     5101 ccaatttgca gcaccacacc gtactgtttg ttacaacagg cataacaata accctgcaac
     5161 aggttgtagg aaaactcagt gaacgaaatc ccctgatctt cacggttgag acgctgctta
     5221 accgcttctt tgttgatcat ctggttaacg gagaagtgtt tgccaatatc gcgcaggaag
     5281 gtcagcacat tcatattgcc gaaccagtca tagttgttcg ccgcgatagc agagttttct
     5341 ccacagtcga aatcgaggaa cggggcaacc tgcttacgga ttttgtccac ccactcctga
     5401 acagtttctt cggtgttcag cttacgctcg gcagctttga agctcgggtc gccaatcaga
     5461 cccgtcgcgc cgcctacaac cgcaaccggc ttgtggcccg cctgctggaa gcgtttcagg
     5521 cataacaatg gaacaagatg ccccaaatgc aagctgtcag cggtaggatc gaagccgcac
     5581 ccgagcgcga tcgggccttg cgccagtcgc tctgctaacg cttcctcgtc cgtcacctgg
     5641 gctaccagcc cccgctcttg caattgttta atcaagttac tgcttgccat ggtggcgcta
     5701 gccagcttgg gtctccctat agtgagtcgt attaatttcg ataagccagt aagcagtggg
     5761 ttctctagtt agccagagag ctctagacca agtgacgatc acagcgatcc acaaacaaga
     5821 accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc cacacccggc gcgtccttat
     5881 ataatcatcg gcgttcaccg ccccacggag atccctccgc agaatcgccg agaagggact
     5941 acttttcctc gcctgttccg ctctctggaa agaaaaccag tgccctagag tcacccaagt
     6001 cccgtcctaa aatgtccttc tgctgatact ggggttctaa ggccgagtct tatgagcagc
     6061 gggccgctgt cctgagcgtc cgggcggaag gatcaggacg ctcgctgcgc ccttcgtctg
     6121 acgtggcagc gctcgccgtg aggagggggg cgcccgcggg aggcgccaaa acccggcgcg
     6181 gaggccttcg aacggccact agcaaaaaat ggagggggac ggattcgaac cgccgaaccc
     6241 aaagggagcg gatttagagt ccgccgcgtt tagccacttc gctacccctc cggtgtctct
     6301 atcactgata gggaacttat aagtctctat cactgatagg gatttcacgt ttatggtgat
     6361 ttcccagaac acatagcgac atgcaaatat taaaaaatgg tgggggaagg attcgaacct
     6421 tcgaagtctg tgacggcaga tttagagtct gctccctttg gccgctcggg aaccccaccg
     6481 gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg
     6541 gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat
     6601 tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt
     6661 ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat
     6721 aggccctctt cctgcccgac ctagcaaaaa atggaggggg acggattcga accgccgaac
     6781 ccaaagggag cggatttaga gtccgccgcg tttagccact tcgctacccc tccggtgtct
     6841 ctatcactga tagggaactt ataagtctct atcactgata gggatttcac gtttatggtg
     6901 atttcccaga acacatagcg acatgcaaat attaaaaaat ggtgggggaa ggattcgaac
     6961 cttcgaagtc tgtgacggca gatttagagt ctgctccctt tggccgctcg ggaaccccac
     7021 cggtgtttcg tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta
     7081 cggtaagcat atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa
     7141 attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa
     7201 ttctaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa
     7261 ataggccctc ttcctgcccg acctagcaaa aaatggaggg ggacggattc gaaccgccga
     7321 acccaaaggg agcggattta gagtccgccg cgtttagcca cttcgctacc cctccggtgt
     7381 ctctatcact gatagggaac ttataagtct ctatcactga tagggatttc acgtttatgg
     7441 tgatttccca gaacacatag cgacatgcaa atattaaaaa atggtggggg aaggattcga
     7501 accttcgaag tctgtgacgg cagatttaga gtctgctccc tttggccgct cgggaacccc
     7561 accggtgttt cgtcctttcc acaagatata taaagccaag aaatcgaaat actttcaagt
     7621 tacggtaagc atatgatagt ccattttaaa acataatttt aaaactgcaa actacccaag
     7681 aaattattac tttctacgtc acgtattttg tactaatatc tttgtgttta cagtcaaatt
     7741 aattctaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag gaatcatggg
     7801 aaataggccc tcttcctgcc cgacctagca aaaaatggag ggggacggat tcgaaccgcc
     7861 gaacccaaag ggagcggatt tagagtccgc cgcgtttagc cacttcgcta cccctccggt
     7921 gtctctatca ctgataggga acttataagt ctctatcact gatagggatt tcacgtttat
     7981 ggtgatttcc cagaacacat agcgacatgc aaatattaaa aaatggtggg ggaaggattc
     8041 gaaccttcga agtctgtgac ggcagattta gagtctgctc cctttggccg ctcgggaacc
     8101 ccaccggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa
     8161 gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca
     8221 agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa
     8281 ttaattctaa ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg
     8341 ggaaataggc cctcttcctg cccgacctag tcaataatca atgtcaacgc gtatatctgg
     8401 cccgtacatc gcgaagcagc gcaaaacgga tcctgcaggc tgatcactgc ttgagcctag
     8461 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     8521 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     8581 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     8641 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     8701 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     8761 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     8821 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     8881 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     8941 ggggcgcgcg ccrggggggg sggggsgggg sggggggsgg ggsggggsgr ggsggrgagg
     9001 kgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     9061 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     9121 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     9181 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     9241 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     9301 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
     9361 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
     9421 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
     9481 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
     9541 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
     9601 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
     9661 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccggggr
     9721 gggctcgggg grggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
     9781 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
     9841 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     9901 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     9961 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
    10021 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
    10081 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
    10141 tgtgctgtct catcattttg gcaaagaatt ggccaaggag gcccatggtg tagagcaagg
    10201 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
    10261 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacttgaggc aagctgaccc
    10321 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
    10381 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
    10441 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
    10501 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
    10561 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
    10621 actacaacag ccacaacgtc taaatcatgg ccgacaagca gaagaacggc atcaaggtga
    10681 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
    10741 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
    10801 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
    10861 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggggccc ttcgaacaaa
    10921 aactcatctc agaagaggat ctgaatatgc ataccggtca tcatcaccat caccatcatc
    10981 acgaaaatct ttattttcaa ggtggaggaa gtggagaaag tttgtttaag gggccgcgtg
    11041 attacaaccc gatatcgagc accatttgtc atttgacgaa tgaatctgat gggcacacaa
    11101 catcgttgta tggtattgga tttggtccct tcatcattac aaacaagcac ttgtttagaa
    11161 gaaataatgg aacactgttg gtccaatcac tacatggtgt attcaaggtc aagaacacca
    11221 cgactttgca acaacacctc attgatggga gggacatgat aattattcgc atgcctaagg
    11281 atttcccacc atttcctcaa aagctgaaat ttagagagcc acaaagggaa gagcgcatat
    11341 gtcttgtgac aaccaacttc caaactaaga gcatgtctag catggtgtca gacactagtt
    11401 gcacattccc ttcatctgat ggcatattct ggaagcattg gattcaaacc aaggatgggc
    11461 agtgtggcag tccattagta tcaactagag atgggttcat tgttggtata cactcagcat
    11521 cgaatttcac caacacaaac aattatttca caagcgtgcc gaaaaacttc atggaattgt
    11581 tgacaaatca ggaggcgcag cagtgggtta gtggttggcg attaaatgct gactcagtat
    11641 tgtggggggg ccataaagtt ttcatggtga aacctgaaga gccttttcag ccagttaagg
    11701 aagcgactca actcatgaat cgtcgtcgcc gtcgctaata ataaggcctc taaggccgaa
    11761 ttcaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
    11821 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
    11881 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
    11941 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
    12001 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
    12061 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
    12121 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
    12181 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
    12241 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
    12301 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
    12361 ctccccgcgt cgactttaac tcggccagca cagtggtcga tcgaccaatg ccctggctca
    12421 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
    12481 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
    12541 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
    12601 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
    12661 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
    12721 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
    12781 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
    12841 actactccca gtcatagctg tccctcttct ttgcggccgc ggtccgtata ctccggaata
    12901 ttaatagatc atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac
    12961 tgttttcgta acagttttgt aataaaaaaa cctataaata ttccggatta ttcataccgt
    13021 cccaccatcg ggcgcgaact cctaaaaaac cgccaccatg aagtgccttt tgtacttagc
    13081 ctttttattc attggggtga attgcaagtt caccatagtt tttccacaca accaaaaagg
    13141 aaactggaaa aatgttcctt ctaattacca ttattgcccg tcaagctcag atttaaattg
    13201 gcataatgac ttaataggca cagccttaca agtcaaaatg cccaagagtc acaaggctat
    13261 tcaagcagac ggttggatgt gtcatgcttc caaatgggtc actacttgtg atttccgctg
    13321 gtatggaccg aagtatataa cacattccat ccgatccttc actccatctg tagaacaatg
    13381 caaggaaagc attgaacaaa cgaaacaagg aacttggctg aatccaggct tccctcctca
    13441 aagttgtgga tatgcaactg tgacggatgc cgaagcagtg attgtccagg tgactcctca
    13501 ccatgtgctg gttgatgaat acacaggaga atgggttgat tcacagttca tcaacggaaa
    13561 atgcagcaat tacatatgcc ccactgtcca taactctaca acctggcatt ctgactataa
    13621 ggtcaaaggg ctatgtgatt ctaacctcat ttccatggac atcaccttct tctcagagga
    13681 cggagagcta tcatccctgg gaaaggaggg cacagggttc agaagtaact actttgctta
    13741 tgaaactgga ggcaaggcct gcaaaatgca atactgcaag cattggggag tcagactccc
    13801 atcaggtgtc tggttcgaga tggctgataa ggatctcttt gctgcagcca gattccctga
    13861 atgcccagaa gggtcaagta tctctgctcc atctcagacc tcagtggatg taagtctaat
    13921 tcaggacgtt gagaggatct tggattattc cctctgccaa gaaacctgga gcaaaatcag
    13981 agcgggtctt ccaatctctc cagtggatct cagctatctt gctcctaaaa acccaggaac
    14041 cggtcctgct ttcaccataa tcaatggtac cctaaaatac tttgagacca gatacatcag
    14101 agtcgatatt gctgctccaa tcctctcaag aatggtcgga atgatcagtg gaactaccac
    14161 agaaagggaa ctgtgggatg actgggcacc atatgaagac gtggaaattg gacccaatgg
    14221 agttctgagg accagttcag gatataagtt tcctttatac atgattggac atggtatgtt
    14281 ggactccgat cttcatctta gctcaaaggc tcaggtgttc gaacatcctc acattcaaga
    14341 cgctgcttcg caacttcctg atgatgagag tttatttttt ggtgatactg ggctatccaa
    14401 aaatccaatc gagcttgtag aaggttggtt cagtagttgg aaaagctcta ttgcctcttt
    14461 tttctttatc atagggttaa tcattggact attcttggtt ctccgagttg gtatccatct
    14521 ttgcattaaa ttaaagcaca ccaagaaaag acagatttat acagacatag agatgaaccg
    14581 acttggaaag tgataaaagc ttgtcgagaa gtactagagg atcataatca gccataccac
    14641 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
    14701 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
    14761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    14821 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag
    14881 gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
    14941 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
    15001 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
    15061 catttttctt
//
