LOCUS       sequence-297114-       10527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with 4xEcoLeuT(CUA) cassette and E. coli AnapRS for amber 
            suppression and incorporation of the fluorescent ncAA Anap; for 
            transient or stable piggyBac-mediated integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10527)
  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 10527)
  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)_AnapRS
FEATURES             Location/Qualifiers
     source          1..10527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     129..151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     143..160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     144..160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   290..352
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     tRNA            complement(join(796..842,843..845,846..879))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     tRNA            complement(join(1144..1190,1191..1193,1194..1227))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     tRNA            complement(join(1491..1537,1538..1540,1541..1574))
                     /product="amber suppressor variant of an E. coli leucine 
                     tRNA (Wu et al., 2004)"
                     /label=tRNA(CUA)
     promoter        1874..2085
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2098..2366
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      2400..2409
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5016..5588
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5183..5200)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5410..5429
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5597..6391
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5651..5670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6261..6280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6451..6471)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6513..6634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6550..6569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6604..6623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7397..7431
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7922..7938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7922..7938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7935..7957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7946..7962
                     /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(7970..7976,7977..7994,7995..8000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8015..8036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8153..8170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8324..8912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8404..8423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9083..9874,9875..9943))
                     /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     9706..9725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9944..10048)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(10074..10527,1..2))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10211..10232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10423..10442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
       61 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      121 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcct cgttcattca
      181 cgtttttgaa cccgtggagg acgggcagac tcgcggtgca aatgtgtttt acagcgtgat
      241 ggagcagatg aagatgctcg acacgctgca gaacacgcag ctagattaac cctagaaaga
      301 taatcatatt gtgacgtacg ttaaagataa tcatgtgtaa aattgacgca tgtgttttat
      361 cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt attatattta
      421 cacttacata ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa
      481 aaaaacaaaa actcaaaatt tcttctataa agtaacaaaa cttttatgag ggacagcccc
      541 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
      601 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
      661 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
      721 tctttgagcc tgcagacacc tggggggata cggggaaaag gcctccacgg ccaactagta
      781 aaaaccgcac ttgtctaccc ggagcgggac ttgaacccgc acagcgcgaa cgccgaggga
      841 tttagaatcc cttgtgtcta ccgattccac catccgggcg aggtacccag gcggcgcaca
      901 agctatataa acctgaagga aatctcaact ttacacttag gtcaagttac ttatcgtact
      961 agagcttcag caggaaattt aactaaaatc taatttaacc agcatagcaa atatcattta
     1021 ttcccaaaat gctaaagttt gagataaacg gacttgattt ccggctgttt tgacactatc
     1081 cagaatgcct tgcagatggg tggggcatgc taaatactgc aggcaagaaa aaccgcactt
     1141 gtctacccgg agcgggactt gaacccgcac agcgcgaacg ccgagggatt tagaatccct
     1201 tgtgtctacc gattccacca tccgggcgag gtacccaggc ggcgcacaag ctatataaac
     1261 ctgaaggaaa tctcaacttt acacttaggt caagttactt atcgtactag agcttcagca
     1321 ggaaatttaa ctaaaatcta atttaaccag catagcaaat atcatttatt cccaaaatgc
     1381 taaagtttga gataaacgga cttgatttcc ggctgttttg acactatcca gaatgccttg
     1441 cagatgggtg gggcatgcta aatactgcag cgataaaaac cgcacttgtc tacccggagc
     1501 gggacttgaa cccgcacagc gcgaacgccg agggatttag aatcccttgt gtctacmgat
     1561 tccaccatcc gggcgaggta cccaggcggc gcacaagcta tataaacctg aaggaaatct
     1621 caactttaca cttaggtcaa gttacttatc gtactagagc ttcagcagga aatttaacta
     1681 aaatctaatt taaccagcat agcaaatatc atttattccc aaaatgctaa agtttgagat
     1741 aaacggactt gatttccggc tgttttgaca ctatccagaa tgccttgcag atgggtgggg
     1801 catgctaaat actgcagctc caaggccact agtgctctag aaaggatctg cgatcgctcc
     1861 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     1921 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     1981 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     2041 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     2101 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     2161 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     2221 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     2281 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     2341 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctactct agagctagcg
     2401 ccaccatgga agagcaatac cggcccgaag agatcgagag caaggtgcag ctgcattggg
     2461 acgagaagcg gaccttcgaa gtgaccgagg acgagagcaa agagaagtac tactgcttca
     2521 gcggccctcc ctaccccagc ggcagactgc acatgggaca cgtgcggaac tacaccatcg
     2581 gcgacgtgat cgccagatac cagcggatgc tgggcaagaa cgtgctgcag cccatcggct
     2641 gggatgcctt tggactgcct gctgaaggcg ccgctgtgaa gaacaacaca gcccctgccc
     2701 cctggaccta cgacaatatc gcctacatga agaaccagct gaagatgctg ggattcggct
     2761 acgactggtc ccgcgagctg gccacctgta cccccgagta ctacagatgg gagcagaagt
     2821 tcttcaccga gctgtacaag aaaggcctgg tgtacaaaaa gaccagcgcc gtgaactggt
     2881 gccccaacga ccagaccgtg ctggccaacg aacaagtgat cgacggctgc tgttggagat
     2941 gcgacaccaa ggtggaacgg aaagaaatcc cccagtggtt catcaagatc accgcctacg
     3001 ccgacgagct gctgaacgac ctggacaagc tggaccactg gcccgacacc gtgaaaacca
     3061 tgcagcggaa ctggatcggc cgcagcgagg gcgtggaaat caccttcaac gtgaacgact
     3121 acgacaacac cctgaccgtg tacaccacca gacccgacac cttcatgggc tgtacctacc
     3181 tggccgtggc cgctggacat cctctggccc agaaagccgc cgagaacaac cctgagctgg
     3241 ccgccttcat cgacgagtgc cggaacacaa aggtggccga ggccgagatg gccacaatgg
     3301 aaaagaaagg cgtggacacc ggcttcaagg ccgtgcaccc tctgaccggc gaagagattc
     3361 ctgtgtgggc cgccaacttc gtgctgatgg aatacggcac aggcgccgtg atggccgtgc
     3421 ctggacacga ccagagagac tacgagttcg cctctaagta cggcctgaac atcaagcccg
     3481 tgatcctggc cgccgatggc agcgagcctg atctgtctca gcaggccctg accgagaagg
     3541 gcgtgctgtt caacagcggc gagttcaacg gcctggacca cgaggccgcc tttaacgcca
     3601 ttgccgacaa gctgaccgcc atgggcgtgg gcgagcggaa agtgaactac cggctgagag
     3661 actggggcgt gtcccggcag agatattggg gagcccccat ccccatggtc accctggaag
     3721 atggcaccgt gatgcccacc cccgacgatc agctgcctgt gatcctgccc gaggacgtcg
     3781 tgatggacgg catcaccagc cccatcaagg ccgatcctga gtgggccaag accaccgtga
     3841 atggcatgcc cgccctgaga gagacagaca ccttcgatac ctttatggaa agcagctggg
     3901 tgctggcccg gtacacctgt ccccagtaca aagaaggcat gctggacagc gaggctgcca
     3961 actactggct gcccgtggat atcgccatcg gaggcatcga gcacgccatc atggaaagcc
     4021 tgtactgccg gttcttccac aagctgatgc gggacgccgg catggtcaat agcgacgagc
     4081 ctgtgaaaca gctgctgtgc cagggcatgg tgctggctga cgccttctac tatgtgggcg
     4141 agaacggcga gagaaactgg gtgtcccccg tggatgccat cgtggaacgg gatgagaagg
     4201 gccggatcgt gaaggccaaa gacgctgccg gacacgagct ggtgtatacc ggcatgagca
     4261 agatgtccaa gagcaagaac aacggcatcg acccccaagt gatggtggaa agatacggcg
     4321 ccgataccgt gcggctgttc atgatgtttg ccagccctgc cgacatgacc ctggaatggc
     4381 aggaatccgg cgtggaaggc gccaaccggt tcctgaagcg cgtgtggaaa ctggtgtacg
     4441 agcacaccgc caagagcgac gtggccgccc tgaatgtgga tgccctgaca gaaaaccaga
     4501 aggccctgcg gcgggacgtg cacaagacaa tcgccaaagt gacagacgac atcggcagac
     4561 ggcagacctt caacaccgct atcgccgcca ttatggaact gatgaacaag ctggccaagg
     4621 cccccaccga cggcgaacag gatagagccc tgatgcagga agccctgctg gctgtcgtgc
     4681 gcatgctgaa ccctttcacc ccccacatct gcttcaccct gtggcaggaa ctgaagggcg
     4741 agggcgacat cgacaacgcc ccttggcctg tggccgacga gaaagccatg gtggaagatt
     4801 ccaccctggt ggtggtgcaa gtgaacggca aagtgcgggc caagatcaca gtgcctgtgg
     4861 acgccaccga ggaacaagtg cgggaaagag ccggccagga acacctggtg gccaagtacc
     4921 tggatggcgt gaccgtgcgc aaagtgatct acgtgcccgg caaactgctg aacctggtcg
     4981 ttggcgggcc cgtttgatga ggatccgcgg ccgcgcccct ctccctcccc cccccctaac
     5041 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     5101 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     5161 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     5221 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     5281 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     5341 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     5401 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     5461 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     5521 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     5581 cgataatacc tccggaatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5641 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5701 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5761 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggccg cgacgggcgt
     5821 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5881 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5941 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6001 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6061 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6121 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6181 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6241 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6301 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6361 cttctatcgc cttcttgacg agttcttctg agtcgacaat caacctctgg attacaaaat
     6421 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6481 tgctttaatg cctttgtatc atgcgttaac taaacttgtt tattgcagct tataatggtt
     6541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggaattga ctcaaatgat
     6661 gtcaattagt ctatcagaag ctcatctggt ctcccttccg ggggacaaga catccctgtt
     6721 taatatttaa acagcagtgt tcccaaactg ggttcttata tcccttgctc tggtcaacca
     6781 ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta aagaaacagt ggcagccagg
     6841 tttagccccg gaattgactg gattcctttt ttagggccca ttggtatggc tttttccccg
     6901 tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc
     6961 ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg
     7021 ggagcgccgg accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg
     7081 acgtaattac atccctgggg gctttggggg ggggctgtcc ctgatatcta taacaagaaa
     7141 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
     7201 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
     7261 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
     7321 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
     7381 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaatctag
     7441 ctgcatcagg atcatatcgt cgggtctttt ttccggctca gtcatcgccc aagctggcgc
     7501 tatctgggca tcggggagga agaagcccgt gccttttccc gcgaggttga agcggcatgg
     7561 aaagagtttg ccgaggatga ctgctgctgc attgacgttg agcgaaaacg cacgtttacc
     7621 atgatgattc gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc
     7681 acctgggata ccagttcgtc gcggcttttc cggacacagt tccggatggt cagcccgaag
     7741 cgcatcagca acccgaacaa taccggcgac agccggaact gccgtgccgg tgtgcagatt
     7801 aatgacagcg gtgcggcgct gggatattac gtcagcgagg acgggtatcc tggctggatg
     7861 ccgcagaaat ggacatggat accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat
     7921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7981 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     8041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     8101 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     8161 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8221 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8281 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8341 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8401 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8461 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8521 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8581 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8641 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8701 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8761 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8821 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8881 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8941 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     9001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     9901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     9961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    10021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
    10081 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
    10141 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
    10201 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
    10261 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
    10321 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
    10381 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
    10441 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
    10501 cgcgcttaat gcgccgctac agggcgc
//
