LOCUS       sequence-459914-       12859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EcTyr aaRS/tRNA system for mammalian ncAA incorporation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12859)
  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 12859)
  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: pAcBac1-EcTyr-TAG
FEATURES             Location/Qualifiers
     source          1..12859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(110..115,116..132,133..138))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    568..792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     781..798
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     regulatory      2143..2152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2171..2190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2172..2190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2243..2642)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     promoter        2671..2919
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        3009..3103
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    3057..3075
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3085..3103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        3213..3461
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        3551..3645
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    3599..3617
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3627..3645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        3755..4003
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        4093..4187
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    4141..4159
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4169..4187
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        4297..4545
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        4635..4729
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    4683..4701
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4711..4729
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        4839..5087
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        5177..5271
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    5225..5243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5253..5271
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5381..5629
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        5719..5813
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    5767..5785
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5795..5813
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5923..6171
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        6261..6355
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    6309..6327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6337..6355
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        6465..6713
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        6803..6897
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    6851..6869
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6879..6897
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7104..7195
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7120..7137
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(7254..7301,7302..8789)
                     /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    8935..9069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8972..8991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9026..9045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9098..9263
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      9447..9901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9534..9553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9743..9764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9927..10031
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10032..10100,10101..10892)
                     /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(10250..10269)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11063..11651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11552..11571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11805..11822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  11956..12180
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(12247..12780)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
       61 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac
      121 cgaacaggct tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc
      181 ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa
      241 ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt
      301 gctgtgcacg gatctgccct tgcttcagga gatcggtaga cctcggccgt cgcggcgctt
      361 gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca
      421 tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctacgta cccgtagtgg
      481 ctatggcagg gcttgcgctt aatgcgccgc tacagggcgc gtggggatac cccctagagc
      541 cccagctggt tctttccgcc tcagaagcca tagagcccac cgcatcccca gcatgcctgc
      601 tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat
      661 gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag
      721 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
      781 tagaaggcac agtcgaggct gatcagcggg tttaaacggg ccctctagac tcgagttaaa
      841 gtcgacttaa cgcgttgaat tcttaaacgg gccctttcca gcaaatcaga cagtaattct
      901 ttttaccgcg acgcagtaag gtaaaacgac caaacagacg atcttcttct ttaaagaagt
      961 attcaggatc ggactgtttt tcaccgttaa tggtgatggc attggaggcg atagttttac
     1021 gtgcctgacc acgggaaggt tgcagttcag aatcgaccag tgcctgcatc aggtctgcgc
     1081 ccttttccat ctcaaccatc ggtacgccgt cctgcgccag ctgttcgaag tccgcttcac
     1141 tcagcgcact caaagaaccg ctgaacaggc attcggtaat acgttttgcc gcctgtaaac
     1201 cttcttcacc gtgaaccaga cgagtcacct gctccgccag tacatactgg gcgcgcggtg
     1261 ctttaccgct gtttttatct tcttcttcca gggcgttgat ctcttcaatg ctcataaagg
     1321 tgaagaactt caggaagcgg taaacgtcgg cacgcgcagt gttgatccag aactggtaga
     1381 atttgtacgg gctggttttc ttcggatcca accagactgc gccgccttca gttttaccaa
     1441 atttggtgcc atctgcttta gtgatcagcg gaacggtcag gccaaacacc tgattctgat
     1501 gcagacgacg ggtcaggtcg ataccagaag tgatgttacc ccactggtca gaaccaccaa
     1561 tttgcagcac cacaccgtac tgtttgttca aacaggccat actataaccc tgcagcaggt
     1621 tgtaggaaaa ctcagtgaac gaaatcccct gatcttcacg gttgagacgc tgcttaaccg
     1681 cttctttgtt gatcatctgg ttaacggaga agtgtttgcc aatatcgcgc aggaaggtca
     1741 gcacattcat attgccgaac cagtcataat tattggccgc gatagcagag ttttctccac
     1801 agtcgaaatc gaggaacggg gcaacctgct tacggatttt gtccacccac tcctgaacag
     1861 tttcttcggt gttcagctta cgctcggcag ctttgaagct cgggtcgcca atcagacccg
     1921 tcgcgccgcc taccagcgca accggcttgt ggcccgcctg ctggaagcgt ttcaggcata
     1981 acaatggaac aagatgcccc aaatgcaagc tgtcagcggt aggatcgaag ccacacacga
     2041 gtgcgatcgg gccttgcgcc agtcgctctg ctaacgcttc ctcgtccgtc acctgggcta
     2101 ccagcccccg ctcttgcaat tgtttaatca agttactgct tgccatggtg gcgctagcca
     2161 gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct
     2221 ctagttagcc agagagctct agaccaagtg acgatcacag cgatccacaa acaagaaccg
     2281 cgacccaaat cccggctgcg acggaactag ctgtgccaca cccggcgcgt ccttatataa
     2341 tcatcggcgt tcaccgcccc acggagatcc ctccgcagaa tcgccgagaa gggactactt
     2401 ttcctcgcct gttccgctct ctggaaagaa aaccagtgcc ctagagtcac ccaagtcccg
     2461 tcctaaaatg tccttctgct gatactgggg ttctaaggcc gagtcttatg agcagcgggc
     2521 cgctgtcctg agcgtccggg cggaaggatc aggacgctcg ctgcgccctt cgtctgacgt
     2581 ggcagcgctc gccgtgagga ggggggcgcc cgcgggaggc gccaaaaccc ggcgcggagg
     2641 ccttcgaacg gccactaggt cgggcaggaa gagggcctat ttcccatgat tccttcatat
     2701 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2761 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2821 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2881 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gtggggttcc cgagcggcca
     2941 aagggagcag actctaaatc tgccgtcaca gacttcgaag gttcgaatcc ttcccccacc
     3001 attttttaat atttgcatgt cgctatgtgt tctgggaaat caccataaac gtgaaatccc
     3061 tatcagtgat agagacttat aagttcccta tcagtgatag agacaccgga ggggtagcga
     3121 agtggctaaa cgcggcggac tctaaatccg ctccctttgg gttcggcggt tcgaatccgt
     3181 ccccctccat tttttgctag gtcgggcagg aagagggcct atttcccatg attccttcat
     3241 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
     3301 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     3361 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     3421 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggtggggtt cccgagcggc
     3481 caaagggagc agactctaaa tctgccgtca cagacttcga aggttcgaat ccttccccca
     3541 ccatttttta atatttgcat gtcgctatgt gttctgggaa atcaccataa acgtgaaatc
     3601 cctatcagtg atagagactt ataagttccc tatcagtgat agagacaccg gaggggtagc
     3661 gaagtggcta aacgcggcgg actctaaatc cgctcccttt gggttcggcg gttcgaatcc
     3721 gtccccctcc attttttgct aggtcgggca ggaagagggc ctatttccca tgattccttc
     3781 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
     3841 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
     3901 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     3961 cgatttcttg gctttatata tcttgtggaa aggacgaaac accggtgggg ttcccgagcg
     4021 gccaaaggga gcagactcta aatctgccgt cacagacttc gaaggttcga atccttcccc
     4081 caccattttt taatatttgc atgtcgctat gtgttctggg aaatcaccat aaacgtgaaa
     4141 tccctatcag tgatagagac ttataagttc cctatcagtg atagagacac cggaggggta
     4201 gcgaagtggc taaacgcggc ggactctaaa tccgctccct ttgggttcgg cggttcgaat
     4261 ccgtccccct ccattttttg ctaggtcggg caggaagagg gcctatttcc catgattcct
     4321 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     4381 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     4441 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     4501 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggtgg ggttcccgag
     4561 cggccaaagg gagcagactc taaatctgcc gtcacagact tcgaaggttc gaatccttcc
     4621 cccaccattt tttaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga
     4681 aatccctatc agtgatagag acttataagt tccctatcag tgatagagac accggagggg
     4741 tagcgaagtg gctaaacgcg gcggactcta aatccgctcc ctttgggttc ggcggttcga
     4801 atccgtcccc ctccattttt tgctaggtcg ggcaggaaga gggcctattt cccatgattc
     4861 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     4921 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     4981 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     5041 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggt ggggttcccg
     5101 agcggccaaa gggagcagac tctaaatctg ccgtcacaga cttcgaaggt tcgaatcctt
     5161 cccccaccat tttttaatat ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt
     5221 gaaatcccta tcagtgatag agacttataa gttccctatc agtgatagag acaccggagg
     5281 ggtagcgaag tggctaaacg cggcggactc taaatccgct ccctttgggt tcggcggttc
     5341 gaatccgtcc ccctccattt tttgctaggt cgggcaggaa gagggcctat ttcccatgat
     5401 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac
     5461 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     5521 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     5581 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg gtggggttcc
     5641 cgagcggcca aagggagcag actctaaatc tgccgtcaca gacttcgaag gttcgaatcc
     5701 ttcccccacc attttttaat atttgcatgt cgctatgtgt tctgggaaat caccataaac
     5761 gtgaaatccc tatcagtgat agagacttat aagttcccta tcagtgatag agacaccgga
     5821 ggggtagcga agtggctaaa cgcggcggac tctaaatccg ctccctttgg gttcggcggt
     5881 tcgaatccgt ccccctccat tttttgctag gtcgggcagg aagagggcct atttcccatg
     5941 attccttcat atttgcatat acgatacaag gctgttagag agataattag aattaatttg
     6001 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     6061 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     6121 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cggtggggtt
     6181 cccgagcggc caaagggagc agactctaaa tctgccgtca cagacttcga aggttcgaat
     6241 ccttccccca ccatttttta atatttgcat gtcgctatgt gttctgggaa atcaccataa
     6301 acgtgaaatc cctatcagtg atagagactt ataagttccc tatcagtgat agagacaccg
     6361 gaggggtagc gaagtggcta aacgcggcgg actctaaatc cgctcccttt gggttcggcg
     6421 gttcgaatcc gtccccctcc attttttgct aggtcgggca ggaagagggc ctatttccca
     6481 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
     6541 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     6601 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     6661 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accggtgggg
     6721 ttcccgagcg gccaaaggga gcagactcta aatctgccgt cacagacttc gaaggttcga
     6781 atccttcccc caccattttt taatatttgc atgtcgctat gtgttctggg aaatcaccat
     6841 aaacgtgaaa tccctatcag tgatagagac ttataagttc cctatcagtg atagagacac
     6901 cggaggggta gcgaagtggc taaacgcggc ggactctaaa tccgctccct ttgggttcgg
     6961 cggttcgaat ccgtccccct ccattttttg ctagtcaata atcaatgtca acgcgtatat
     7021 ctggcccgta catcgcgaag cagcgcaaaa cggatcctgc aggtatttgc ggccgcggtc
     7081 cgtatactcc ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa
     7141 taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc
     7201 ggattattca taccgtccca ccatcgggcg cgaactccta aaaaaccgcc accatgaagt
     7261 gccttttgta cttagccttt ttattcattg gggtgaattg caagttcacc atagtttttc
     7321 cacacaacca aaaaggaaac tggaaaaatg ttccttctaa ttaccattat tgcccgtcaa
     7381 gctcagattt aaattggcat aatgacttaa taggcacagc cttacaagtc aaaatgccca
     7441 agagtcacaa ggctattcaa gcagacggtt ggatgtgtca tgcttccaaa tgggtcacta
     7501 cttgtgattt ccgctggtat ggaccgaagt atataacaca ttccatccga tccttcactc
     7561 catctgtaga acaatgcaag gaaagcattg aacaaacgaa acaaggaact tggctgaatc
     7621 caggcttccc tcctcaaagt tgtggatatg caactgtgac ggatgccgaa gcagtgattg
     7681 tccaggtgac tcctcaccat gtgctggttg atgaatacac aggagaatgg gttgattcac
     7741 agttcatcaa cggaaaatgc agcaattaca tatgccccac tgtccataac tctacaacct
     7801 ggcattctga ctataaggtc aaagggctat gtgattctaa cctcatttcc atggacatca
     7861 ccttcttctc agaggacgga gagctatcat ccctgggaaa ggagggcaca gggttcagaa
     7921 gtaactactt tgcttatgaa actggaggca aggcctgcaa aatgcaatac tgcaagcatt
     7981 ggggagtcag actcccatca ggtgtctggt tcgagatggc tgataaggat ctctttgctg
     8041 cagccagatt ccctgaatgc ccagaagggt caagtatctc tgctccatct cagacctcag
     8101 tggatgtaag tctaattcag gacgttgaga ggatcttgga ttattccctc tgccaagaaa
     8161 cctggagcaa aatcagagcg ggtcttccaa tctctccagt ggatctcagc tatcttgctc
     8221 ctaaaaaccc aggaaccggt cctgctttca ccataatcaa tggtacccta aaatactttg
     8281 agaccagata catcagagtc gatattgctg ctccaatcct ctcaagaatg gtcggaatga
     8341 tcagtggaac taccacagaa agggaactgt gggatgactg ggcaccatat gaagacgtgg
     8401 aaattggacc caatggagtt ctgaggacca gttcaggata taagtttcct ttatacatga
     8461 ttggacatgg tatgttggac tccgatcttc atcttagctc aaaggctcag gtgttcgaac
     8521 atcctcacat tcaagacgct gcttcgcaac ttcctgatga tgagagttta ttttttggtg
     8581 atactgggct atccaaaaat ccaatcgagc ttgtagaagg ttggttcagt agttggaaaa
     8641 gctctattgc ctcttttttc tttatcatag ggttaatcat tggactattc ttggttctcc
     8701 gagttggtat ccatctttgc attaaattaa agcacaccaa gaaaagacag atttatacag
     8761 acatagagat gaaccgactt ggaaagtgat aaggccaggc cggccaagct tgtcgagaag
     8821 tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     8881 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     8941 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     9001 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     9061 gtctggatct gatcactgct tgagcctagg agatccgaac cagataagtg aaatctagtt
     9121 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
     9181 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
     9241 cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa
     9301 catcctgcca actccatgtg acaaaccgtc atcttcggct actttttctc tgtcacagaa
     9361 tgaaaatttt tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct
     9421 tatttgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     9481 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     9541 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     9601 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     9661 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     9721 gttggagtcc acgttcttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     9781 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9841 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     9901 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     9961 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
    10021 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
    10081 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
    10141 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
    10201 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
    10261 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
    10321 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
    10381 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
    10441 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
    10501 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
    10561 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
    10621 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
    10681 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
    10741 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
    10801 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
    10861 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
    10921 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
    10981 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
    11041 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
    11101 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
    11161 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
    11221 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
    11281 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
    11341 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
    11401 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
    11461 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
    11521 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
    11581 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
    11641 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
    11701 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
    11761 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
    11821 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
    11881 accgcataga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa taaatggatg
    11941 ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa
    12001 ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat
    12061 gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag
    12121 tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg gcatggtaaa gactatattc
    12181 gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac
    12241 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
    12301 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
    12361 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
    12421 ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg
    12481 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
    12541 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
    12601 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
    12661 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
    12721 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
    12781 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
    12841 cccgaggcat agactgtac
//
