LOCUS       sequence-344138-       10696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AnapRS expression for amber suppression in mammalian cells; 
            transient or piggy bac mediated integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10696)
  AUTHORS   Meineke B, Heimgartner J, Caridha R, Block MF, Kimler KJ, Pires MF, 
            Landreh M, Elsasser SJ
  TITLE     Dual stop codon suppression in mammalian cells with genomically 
            integrated genetic code expansion machinery.
  JOURNAL   Cell Rep Methods. 2023 Nov 20;3(11):100626. doi: 
            10.1016/j.crmeth.2023.100626. Epub 2023 Nov 6.
  PUBMED    37935196
REFERENCE   2  (bases 1 to 10696)
  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: "Cell Rep 
            Methods."; date: "2023-11-20"; volume: "3(11):100626. doi"; pages: "
            10.1016/j.crmeth.2023.100626. Epub 2023 Nov 6"
COMMENT     Sequence Label: pAS_4xEcoLeuT(CUA)_EF1_AnapRS
FEATURES             Location/Qualifiers
     source          1..10696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     tRNA            complement(join(1380..1426,1427..1429,1430..1463))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     tRNA            complement(join(1727..1773,1774..1776,1777..1810))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     tRNA            complement(join(2074..2120,2121..2123,2124..2157))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     tRNA            complement(join(2421..2467,2468..2470,2471..2504))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     promoter        2804..3015
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3028..3296
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      3330..3339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(5946..6520,6521..6523,6524..6532)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6113..6130)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6340..6359
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6545..7144
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6545..6564)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7041..7061
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7204..7224)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    7266..7387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7303..7322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7357..7376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8150..8184
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8675..8691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8675..8691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8688..8710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8699..8715
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8723..8729,8730..8747,8748..8753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8768..8789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8906..8923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9077..9665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9157..9176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9836..10627,10628..10696))
                     /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     10459..10478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccaact agtaaaaacc gcacttgtct
     1381 acccggagcg ggacttgaac ccgcacagcg cgaacgccga gggatttaga atcccttgtg
     1441 tctaccgatt ccaccatccg ggcgaggtac ccaggcggcg cacaagctat ataaacctga
     1501 aggaaatctc aactttacac ttaggtcaag ttacttatcg tactagagct tcagcaggaa
     1561 atttaactaa aatctaattt aaccagcata gcaaatatca tttattccca aaatgctaaa
     1621 gtttgagata aacggacttg atttccggct gttttgacac tatccagaat gccttgcaga
     1681 tgggtggggc atgctaaata ctgcaggcaa aaaaaccgca cttgtctacc cggagcggga
     1741 cttgaacccg cacagcgcga acgccgaggg atttagaatc ccttgtgtct accgattcca
     1801 ccatccgggc gaggtaccca ggcggcgcac aagctatata aacctgaagg aaatctcaac
     1861 tttacactta ggtcaagtta cttatcgtac tagagcttca gcaggaaatt taactaaaat
     1921 ctaatttaac cagcatagca aatatcattt attcccaaaa tgctaaagtt tgagataaac
     1981 ggacttgatt tccggctgtt ttgacactat ccagaatgcc ttgcagatgg gtggggcatg
     2041 ctaaatactg cagcaagaaa aaccgcactt gtctacccgg agcgggactt gaacccgcac
     2101 agcgcgaacg ccgagggatt tagaatccct tgtgtctacc gattccacca tccgggcgag
     2161 gtacccaggc ggcgcacaag ctatataaac ctgaaggaaa tctcaacttt acacttaggt
     2221 caagttactt atcgtactag agcttcagca ggaaatttaa ctaaaatcta atttaaccag
     2281 catagcaaat atcatttatt cccaaaatgc taaagtttga gataaacgga cttgatttcc
     2341 ggctgttttg acactatcca gaatgccttg cagatgggtg gggcatgcta aatactgcag
     2401 cgataaaaac cgcacttgtc tacccggagc gggacttgaa cccgcacagc gcgaacgccg
     2461 agggatttag aatcccttgt gtctacagat tccaccatcc gggcgaggta cccaggcggc
     2521 gcacaagcta tataaacctg aaggaaatct caactttaca cttaggtcaa gttacttatc
     2581 gtactagagc ttcagcagga aatttaacta aaatctaatt taaccagcat agcaaatatc
     2641 atttattccc aaaatgctaa agtttgagat aaacggactt gatttccggc tgttttgaca
     2701 ctatccagaa tgccttgcag atgggtgggg catgctaaat actgcagctc caaggccact
     2761 agtgctctag aaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     2821 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     2881 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     2941 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     3001 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     3061 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     3121 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     3181 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     3241 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     3301 ctgtgaccgg cgcctactct agagctagcg ccaccatgga agagcaatac cggcccgaag
     3361 agatcgagag caaggtgcag ctgcattggg acgagaagcg gaccttcgaa gtgaccgagg
     3421 acgagagcaa agagaagtac tactgcttca gcggccctcc ctaccccagc ggcagactgc
     3481 acatgggaca cgtgcggaac tacaccatcg gcgacgtgat cgccagatac cagcggatgc
     3541 tgggcaagaa cgtgctgcag cccatcggct gggatgcctt tggactgcct gctgaaggcg
     3601 ccgctgtgaa gaacaacaca gcccctgccc cctggaccta cgacaatatc gcctacatga
     3661 agaaccagct gaagatgctg ggattcggct acgactggtc ccgcgagctg gccacctgta
     3721 cccccgagta ctacagatgg gagcagaagt tcttcaccga gctgtacaag aaaggcctgg
     3781 tgtacaaaaa gaccagcgcc gtgaactggt gccccaacga ccagaccgtg ctggccaacg
     3841 aacaagtgat cgacggctgc tgttggagat gcgacaccaa ggtggaacgg aaagaaatcc
     3901 cccagtggtt catcaagatc accgcctacg ccgacgagct gctgaacgac ctggacaagc
     3961 tggaccactg gcccgacacc gtgaaaacca tgcagcggaa ctggatcggc cgcagcgagg
     4021 gcgtggaaat caccttcaac gtgaacgact acgacaacac cctgaccgtg tacaccacca
     4081 gacccgacac cttcatgggc tgtacctacc tggccgtggc cgctggacat cctctggccc
     4141 agaaagccgc cgagaacaac cctgagctgg ccgccttcat cgacgagtgc cggaacacaa
     4201 aggtggccga ggccgagatg gccacaatgg aaaagaaagg cgtggacacc ggcttcaagg
     4261 ccgtgcaccc tctgaccggc gaagagattc ctgtgtgggc cgccaacttc gtgctgatgg
     4321 aatacggcac aggcgccgtg atggccgtgc ctggacacga ccagagagac tacgagttcg
     4381 cctctaagta cggcctgaac atcaagcccg tgatcctggc cgccgatggc agcgagcctg
     4441 atctgtctca gcaggccctg accgagaagg gcgtgctgtt caacagcggc gagttcaacg
     4501 gcctggacca cgaggccgcc tttaacgcca ttgccgacaa gctgaccgcc atgggcgtgg
     4561 gcgagcggaa agtgaactac cggctgagag actggggcgt gtcccggcag agatattggg
     4621 gagcccccat ccccatggtc accctggaag atggcaccgt gatgcccacc cccgacgatc
     4681 agctgcctgt gatcctgccc gaggacgtcg tgatggacgg catcaccagc cccatcaagg
     4741 ccgatcctga gtgggccaag accaccgtga atggcatgcc cgccctgaga gagacagaca
     4801 ccttcgatac ctttatggaa agcagctggg tgctggcccg gtacacctgt ccccagtaca
     4861 aagaaggcat gctggacagc gaggctgcca actactggct gcccgtggat atcgccatcg
     4921 gaggcatcga gcacgccatc atggaaagcc tgtactgccg gttcttccac aagctgatgc
     4981 gggacgccgg catggtcaat agcgacgagc ctgtgaaaca gctgctgtgc cagggcatgg
     5041 tgctggctga cgccttctac tatgtgggcg agaacggcga gagaaactgg gtgtcccccg
     5101 tggatgccat cgtggaacgg gatgagaagg gccggatcgt gaaggccaaa gacgctgccg
     5161 gacacgagct ggtgtatacc ggcatgagca agatgtccaa gagcaagaac aacggcatcg
     5221 acccccaagt gatggtggaa agatacggcg ccgataccgt gcggctgttc atgatgtttg
     5281 ccagccctgc cgacatgacc ctggaatggc aggaatccgg cgtggaaggc gccaaccggt
     5341 tcctgaagcg cgtgtggaaa ctggtgtacg agcacaccgc caagagcgac gtggccgccc
     5401 tgaatgtgga tgccctgaca gaaaaccaga aggccctgcg gcgggacgtg cacaagacaa
     5461 tcgccaaagt gacagacgac atcggcagac ggcagacctt caacaccgct atcgccgcca
     5521 ttatggaact gatgaacaag ctggccaagg cccccaccga cggcgaacag gatagagccc
     5581 tgatgcagga agccctgctg gctgtcgtgc gcatgctgaa ccctttcacc ccccacatct
     5641 gcttcaccct gtggcaggaa ctgaagggcg agggcgacat cgacaacgcc ccttggcctg
     5701 tggccgacga gaaagccatg gtggaagatt ccaccctggt ggtggtgcaa gtgaacggca
     5761 aagtgcgggc caagatcaca gtgcctgtgg acgccaccga ggaacaagtg cgggaaagag
     5821 ccggccagga acacctggtg gccaagtacc tggatggcgt gaccgtgcgc aaagtgatct
     5881 acgtgcccgg caaactgctg aacctggtcg ttggcgggcc cgtttgatga ggatccgcgg
     5941 ccgcgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     6001 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     6061 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     6121 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     6181 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     6241 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     6301 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     6361 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     6421 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     6481 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggcgtc
     6541 cggaatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     6601 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     6661 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     6721 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     6781 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     6841 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     6901 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     6961 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     7021 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     7081 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     7141 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     7201 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgcgtt
     7261 aactaaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7321 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7381 tatcttatca tgtctggaat tgactcaaat gatgtcaatt agtctatcag aagctcatct
     7441 ggtctccctt ccgggggaca agacatccct gtttaatatt taaacagcag tgttcccaaa
     7501 ctgggttctt atatcccttg ctctggtcaa ccaggttgca gggtttcctg tcctcacagg
     7561 aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc ccggaattga ctggattcct
     7621 tttttagggc ccattggtat ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     7681 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     7741 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     7801 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     7861 gggggggctg tccctgatat ctataacaag aaaatatata tataataagt tatcacgtaa
     7921 gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa
     7981 gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac
     8041 cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac
     8101 agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat
     8161 tttacgcaga ctatctttct agggttaatc tagctgcatc aggatcatat cgtcgggtct
     8221 tttttccggc tcagtcatcg cccaagctgg cgctatctgg gcatcgggga ggaagaagcc
     8281 cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt ttgccgagga tgactgctgc
     8341 tgcattgacg ttgagcgaaa acgcacgttt accatgatga ttcgggaagg tgtggccatg
     8401 cacgccttta acggtgaact gttcgttcag gccacctggg ataccagttc gtcgcggctt
     8461 ttccggacac agttccggat ggtcagcccg aagcgcatca gcaacccgaa caataccggc
     8521 gacagccgga actgccgtgc cggtgtgcag attaatgaca gcggtgcggc gctgggatat
     8581 tacgtcagcg aggacgggta tcctggctgg atgccgcaga aatggacatg gataccccgt
     8641 gagttacccg gcgggcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     8701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     8761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     8821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     8881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     8941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     9001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     9061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     9121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     9421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     9481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     9541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     9601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     9661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     9721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     9781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     9841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     9901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     9961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    10021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    10081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    10141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    10201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    10441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    10501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    10561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    10621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    10681 aatgttgaat actcat
//
