LOCUS       sequence-455605-       11839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pyl aaRS/tRNA system for mammalian TAG suppression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11839)
  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 11839)
  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-Pyl-TAG
FEATURES             Location/Qualifiers
     source          1..11839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  894..1118
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1185..1718)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1907..1912,1913..1929,1930..1935))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    2365..2589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2578..2595
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    2644..3232
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2750..2761
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3159..3179
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     regulatory      4511..4520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4539..4558)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4540..4558)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4603..4806)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4632..4652)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4807..5110
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(5256..5470)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(5557..5805)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(5906..6120)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(6207..6455)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(6556..6770)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(6857..7105)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(7206..7420)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(7507..7755)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        7881..7972
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7897..7914
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(8031..8078,8079..9566)
                     /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    9712..9846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9749..9768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9803..9822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9875..10040
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      10224..10678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10311..10330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10520..10541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10704..10808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10809..10877,10878..11669)
                     /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(11027..11046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa
      841 cctggcaaaa tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc
      901 ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac
      961 tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac
     1021 ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa
     1081 agaggggcgt ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg
     1141 aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt
     1201 gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact
     1261 gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc
     1321 tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag
     1381 actgcgagat catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa
     1441 gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta
     1501 ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg
     1561 tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg
     1621 ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga
     1681 ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa
     1741 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa
     1801 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     1861 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga
     1921 acaggcttat gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     1981 aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt
     2041 ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct
     2101 gtgcacggat ctgcccttgc ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc
     2161 ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg
     2221 tttgttcgcc caggactcta gctatagttc tagtggttgg ctacgtaccc gtagtggcta
     2281 tggcagggct tgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc
     2341 agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca tgcctgctat
     2401 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
     2461 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
     2521 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
     2581 aaggcacagt cgaggctgat cagcgggttt aaacgggccc tctagactcg agttaaagtc
     2641 gacgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
     2701 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
     2761 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc
     2821 aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
     2881 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
     2941 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
     3001 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
     3061 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     3121 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     3181 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttgtcgactt
     3241 aacgcgttga attcttacag attggttgaa atcccattat agtaagattc ggaccttgat
     3301 gccctcttaa tgtttttaaa gccgtgcata accttgagca agcgttcaag accaaaacct
     3361 gcacctatcc atggtttgtc aataccccat tctctatcaa gagaaactgg cccgacgact
     3421 gccgaagaaa gctccaggtc cccgtgcatt atatcaagag tatccccaaa gaccatacag
     3481 gaatctccta cgatttcgaa gtcgatttcc agatagtcca gaaactcttt gatgagagct
     3541 tcaagatttt cccgagtaca tcccgaaccc atctgacaga agttcaccat agtaaattct
     3601 tccaggtgct ctttgccgtc agactctttc cggtaacaag gtccgacttc gaaaattttt
     3661 attgggcctg gtaaaatcct atcgagtttt cgcagatagt tgtaaagagt cggggcaagc
     3721 attggcctca agcagagatt tttatccacc cggaagatct gttttgaaag ttcagtatca
     3781 ttattaatac ccattctctc cacgtattcc gccggaataa ggataggaga ctttatctcc
     3841 agaaaacccc ggtctacgaa aaatttcgta atatcacgtt cgagtttacc gaggtagtct
     3901 tctctatcat tggtatagag ccgctgaaaa tcgttttttc ttcttgtcac aagttcaggc
     3961 tcaagttccc tgaaaggctt tgccatattt agagaaattt tatcctctgg acttaagaga
     4021 gcctcaaccc tatcaagctg gcttcttgta agtgaaggag caggagccga tgcgggaaca
     4081 gacgaatttg gagttgattt tgcaggcgaa ggtacagatc tggatgtgtt cgttgatgcc
     4141 tttgcagaaa cagaattttc cagaggcttc ggagcccttg aaactgattt cggcatagct
     4201 tttttgacct ttggagcaga aactacccta actttcacac tgtttttgct ttcggttgat
     4261 cttgtgagaa aattattgat atcctcgtcc gaaaccctac atcgtttgca ggtttttctg
     4321 tacttatgat gtctgaatgc tctggctgtt ctacaactcc tggaattatt cacaacaaga
     4381 tggtctccac acgccatttc aatgtatatt ttacttcttg agacctcatg gtgcttgatt
     4441 ttgtggagcg tgccagtcct ggacatccag agcccggtcg cagatattaa aacatctaat
     4501 ggttttttat ccatggtggc gctagccagc ttgggtctcc ctatagtgag tcgtattaat
     4561 ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac
     4621 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     4681 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     4741 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     4801 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     4861 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     4921 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     4981 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     5041 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     5101 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     5161 ttaataacta gcaaaaaacg gaaaccccgg gaatctaacc cggctgaacg gatttagagt
     5221 ccgttcgatc tacatgatca ggtttccggg gatctgtggt ctcatacaga acttataaga
     5281 ttcccaaatc caaagacatt tcacgtttat ggtgatttcc cagaacacat agcgacatgc
     5341 aaatattgca gggcgccact cccctgtccc tcacagccat cttcctgcca gggcgcacgc
     5401 gcgctgggtg ttcccgccta gtgacactgg gcccgcgatt ccttggagcg ggttgatgac
     5461 gtcagcgttc gaattcctag caaaaaacgg aaaccccggg aatctaaccc ggctgaacgg
     5521 atttagagtc cgttcgatct acatgatcag gtttccggtg tttcgtcctt tccacaagat
     5581 atataaagcc aagaaatcga aatactttca agttacggta agcatatgat agtccatttt
     5641 aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt
     5701 ttgtactaat atctttgtgt ttacagtcaa attaattcta attatctctc taacagcctt
     5761 gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcttcct gcccgaccta
     5821 gcaaaaaacg gaaaccccgg gaatctaacc cggctgaacg gatttagagt ccgttcgatc
     5881 tacatgatca ggtttccggg gatctgtggt ctcatacaga acttataaga ttcccaaatc
     5941 caaagacatt tcacgtttat ggtgatttcc cagaacacat agcgacatgc aaatattgca
     6001 gggcgccact cccctgtccc tcacagccat cttcctgcca gggcgcacgc gcgctgggtg
     6061 ttcccgccta gtgacactgg gcccgcgatt ccttggagcg ggttgatgac gtcagcgttc
     6121 gaattcctag caaaaaacgg aaaccccggg aatctaaccc ggctgaacgg atttagagtc
     6181 cgttcgatct acatgatcag gtttccggtg tttcgtcctt tccacaagat atataaagcc
     6241 aagaaatcga aatactttca agttacggta agcatatgat agtccatttt aaaacataat
     6301 tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat
     6361 atctttgtgt ttacagtcaa attaattcta attatctctc taacagcctt gtatcgtata
     6421 tgcaaatatg aaggaatcat gggaaatagg ccctcttcct gcccgaccta gcaaaaaacg
     6481 gaaaccccgg gaatctaacc cggctgaacg gatttagagt ccgttcgatc tacatgatca
     6541 ggtttccggg gatctgtggt ctcatacaga acttataaga ttcccaaatc caaagacatt
     6601 tcacgtttat ggtgatttcc cagaacacat agcgacatgc aaatattgca gggcgccact
     6661 cccctgtccc tcacagccat cttcctgcca gggcgcacgc gcgctgggtg ttcccgccta
     6721 gtgacactgg gcccgcgatt ccttggagcg ggttgatgac gtcagcgttc gaattcctag
     6781 caaaaaacgg aaaccccggg aatctaaccc ggctgaacgg atttagagtc cgttcgatct
     6841 acatgatcag gtttccggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga
     6901 aatactttca agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg
     6961 caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt
     7021 ttacagtcaa attaattcta attatctctc taacagcctt gtatcgtata tgcaaatatg
     7081 aaggaatcat gggaaatagg ccctcttcct gcccgaccta gcaaaaaacg gaaaccccgg
     7141 gaatctaacc cggctgaacg gatttagagt ccgttcgatc tacatgatca ggtttccggg
     7201 gatctgtggt ctcatacaga acttataaga ttcccaaatc caaagacatt tcacgtttat
     7261 ggtgatttcc cagaacacat agcgacatgc aaatattgca gggcgccact cccctgtccc
     7321 tcacagccat cttcctgcca gggcgcacgc gcgctgggtg ttcccgccta gtgacactgg
     7381 gcccgcgatt ccttggagcg ggttgatgac gtcagcgttc gaattcctag caaaaaacgg
     7441 aaaccccggg aatctaaccc ggctgaacgg atttagagtc cgttcgatct acatgatcag
     7501 gtttccggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca
     7561 agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc
     7621 aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa
     7681 attaattcta attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat
     7741 gggaaatagg ccctcttcct gcccgaccta gtcaataatc aatgtcaacg cgtatatctg
     7801 gcccgtacat cgcgaagcag cgcaaaacgg atcctgcagg tatttgcggc cgcggtccgt
     7861 atactccgga atattaatag atcatggaga taattaaaat gataaccatc tcgcaaataa
     7921 ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa atattccgga
     7981 ttattcatac cgtcccacca tcgggcgcga actcctaaaa aaccgccacc atgaagtgcc
     8041 ttttgtactt agccttttta ttcattgggg tgaattgcaa gttcaccata gtttttccac
     8101 acaaccaaaa aggaaactgg aaaaatgttc cttctaatta ccattattgc ccgtcaagct
     8161 cagatttaaa ttggcataat gacttaatag gcacagcctt acaagtcaaa atgcccaaga
     8221 gtcacaaggc tattcaagca gacggttgga tgtgtcatgc ttccaaatgg gtcactactt
     8281 gtgatttccg ctggtatgga ccgaagtata taacacattc catccgatcc ttcactccat
     8341 ctgtagaaca atgcaaggaa agcattgaac aaacgaaaca aggaacttgg ctgaatccag
     8401 gcttccctcc tcaaagttgt ggatatgcaa ctgtgacgga tgccgaagca gtgattgtcc
     8461 aggtgactcc tcaccatgtg ctggttgatg aatacacagg agaatgggtt gattcacagt
     8521 tcatcaacgg aaaatgcagc aattacatat gccccactgt ccataactct acaacctggc
     8581 attctgacta taaggtcaaa gggctatgtg attctaacct catttccatg gacatcacct
     8641 tcttctcaga ggacggagag ctatcatccc tgggaaagga gggcacaggg ttcagaagta
     8701 actactttgc ttatgaaact ggaggcaagg cctgcaaaat gcaatactgc aagcattggg
     8761 gagtcagact cccatcaggt gtctggttcg agatggctga taaggatctc tttgctgcag
     8821 ccagattccc tgaatgccca gaagggtcaa gtatctctgc tccatctcag acctcagtgg
     8881 atgtaagtct aattcaggac gttgagagga tcttggatta ttccctctgc caagaaacct
     8941 ggagcaaaat cagagcgggt cttccaatct ctccagtgga tctcagctat cttgctccta
     9001 aaaacccagg aaccggtcct gctttcacca taatcaatgg taccctaaaa tactttgaga
     9061 ccagatacat cagagtcgat attgctgctc caatcctctc aagaatggtc ggaatgatca
     9121 gtggaactac cacagaaagg gaactgtggg atgactgggc accatatgaa gacgtggaaa
     9181 ttggacccaa tggagttctg aggaccagtt caggatataa gtttccttta tacatgattg
     9241 gacatggtat gttggactcc gatcttcatc ttagctcaaa ggctcaggtg ttcgaacatc
     9301 ctcacattca agacgctgct tcgcaacttc ctgatgatga gagtttattt tttggtgata
     9361 ctgggctatc caaaaatcca atcgagcttg tagaaggttg gttcagtagt tggaaaagct
     9421 ctattgcctc ttttttcttt atcatagggt taatcattgg actattcttg gttctccgag
     9481 ttggtatcca tctttgcatt aaattaaagc acaccaagaa aagacagatt tatacagaca
     9541 tagagatgaa ccgacttgga aagtgataag gccaggccgg ccaagcttgt cgagaagtac
     9601 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     9661 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     9721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     9781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     9841 tggatctgat cactgcttga gcctaggaga tccgaaccag ataagtgaaa tctagttcca
     9901 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     9961 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
    10021 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat
    10081 cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga
    10141 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
    10201 ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    10261 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    10321 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    10381 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    10441 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    10501 ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
    10561 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
    10621 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
    10681 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
    10741 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
    10801 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
    10861 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
    10921 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
    10981 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
    11041 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
    11101 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
    11161 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
    11221 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
    11281 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
    11341 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
    11401 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
    11461 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
    11521 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
    11581 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
    11641 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    11701 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    11761 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    11821 aaaagatcaa aggatcttc
//
