LOCUS       sequence-469600-       10409 bp    DNA     circular SYN 22-APR-2025
DEFINITION  for PiggyBac mediated integration and stable expression of hFGFR1b 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10409)
  AUTHORS   Zlinska V, Feketova Z, Czyrek A, Chudzian J, Zivkovic ML, Ursachi 
            VC, Dudeja P, Fafilek B, Rynes J, Rico-Llanos G, Koudelka A, Roy T, 
            Biadun M, Raskova V, Svozilova K, Stroblova M, Krzyscik M, Hristova 
            K, Krowarsch D, Foldynova-Trantirkova S, Zakrzewska M, Trantirek L, 
            Krejci P
  TITLE     Specific inhibition of fibroblast growth factor receptor 1 signaling
            by a DNA aptamer.
  JOURNAL   Mol Ther Nucleic Acids. 2024 Nov 28;36(1):102405. doi: 
            10.1016/j.omtn.2024.102405. eCollection 2025 Mar 11.
  PUBMED    39759879
REFERENCE   2  (bases 1 to 10409)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Nucleic Acids."; date: "2024-11-28"; volume: "36(1):102405. doi"; 
            pages: " 10.1016/j.omtn.2024.102405. eCollection 2025 Mar 11"
COMMENT     Sequence Label: pTR01F/hFGFR1b-V5
FEATURES             Location/Qualifiers
     source          1..10409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /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(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1521..1540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1774..1791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1908..1929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1944..1949,1950..1967,1968..1974)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1982..1998
                     /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)."
     primer_bind     1987..2009
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2006..2022
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2006..2022
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2513..2547
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     3130..3149
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3283..3486
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3437..3457
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3531..3550
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3531..3549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3986..4003)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6148..6189
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6199..6216
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6267..6388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        6636..6847
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             6860..7128
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      7157..7166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7187..7849
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     CDS             7919..7972
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7973..8572
                     /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(7973..7992)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8469..8489
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8626..8646)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    8688..8809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   9474..9536
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9666..9683)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9666..9682)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9675..9697)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9824..10279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9911..9930)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10121..10142
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10305..10409
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1741 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1801 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     1861 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     1921 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1981 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     2041 gcccgccggg taactcacgg ggtatccatg tccatttctg cggcatccag ccaggatacc
     2101 cgtcctcgct gacgtaatat cccagcgccg caccgctgtc attaatctgc acaccggcac
     2161 ggcagttccg gctgtcgccg gtattgttcg ggttgctgat gcgcttcggg ctgaccatcc
     2221 ggaactgtgt ccggaaaagc cgcgacgaac tggtatccca ggtggcctga acgaacagtt
     2281 caccgttaaa ggcgtgcatg gccacacctt cccgaatcat catggtaaac gtgcgttttc
     2341 gctcaacgtc aatgcagcag cagtcatcct cggcaaactc tttccatgcc gcttcaacct
     2401 cgcgggaaaa ggcacgggct tcttcctccc cgatgcccag atagcgccag cttgggcgat
     2461 gactgagccg gaaaaaagac ccgacgatat gatcctgatg cagctagatt aaccctagaa
     2521 agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata
     2581 gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg
     2641 cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa
     2701 atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg
     2761 ttatttcatg ttctacttac gtgataactt attatatata tattttcttg ttatagatat
     2821 cagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
     2881 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
     2941 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
     3001 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaggcctcca
     3061 cggccactag tattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3121 ctacgtaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     3181 tggcctaact ggccggtacc tgagctcgct agcgggaatt tccggggact ttccgggaat
     3241 ttccggggac tttccgggaa tttccagatc gctattacca tggtgatgcg gttttggcag
     3301 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     3361 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     3421 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     3481 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     3541 cactataggg agacccaagc tggctagtta agcttggtac cgagctcgga tccagtaccc
     3601 ttcaccatgt ggagatggaa gtgcctcctc ttctgggctg tgctggtcac agccacactc
     3661 tgcaccgcta ggccgtcccc gaccttgcct gaacaagccc agccctgggg agcccctgtg
     3721 gaagtggagt ccttcctggt ccaccccggt gacctgctgc agcttcgctg tcggctgcgg
     3781 gacgatgtgc agagcatcaa ctggctgcgg gacggggtgc agctggcgga aagcaaccgc
     3841 acccgcatca caggggagga ggtggaggtg caggactccg tgcccgcaga ctccggcctc
     3901 tatgcttgcg taaccagcag cccctcgggc agtgacacca cctacttctc cgtcaatgtt
     3961 tcagatgctc tcccctcctc ggaggatgat gatgatgatg atgactcctc ttcagaggag
     4021 aaagaaacag ataacaccaa accaaacccc gtagctccat attggacatc cccagaaaag
     4081 atggaaaaga aattgcatgc agtgccggct gccaagacag tgaagttcaa atgcccttcc
     4141 agtgggaccc caaaccccac actgcgctgg ttgaaaaatg gcaaagaatt caaacctgac
     4201 cacagaattg gaggctacaa ggtccgttat gccacctgga gcatcataat ggactctgtg
     4261 gtgccctctg acaagggcaa ctacacctgc attgtggaga atgagtacgg cagcatcaac
     4321 cacacatacc agctggatgt cgtggagcgg tcccctcacc ggcccatcct gcaagcaggg
     4381 ttgcccgcca acaaaacagt ggccctgggt agcaacgtgg agttcatgtg taaggtgtac
     4441 agtgacccgc agccgcacat ccagtggcta aagcacatcg aggtgaatgg gagcaagatt
     4501 ggcccagaca acctgcctta tgtccagatc ttgaagcatt cggggattaa tagctcggat
     4561 gcggaggtgc tgaccctgtt caatgtgaca gaggcccaga gcggggagta tgtgtgtaag
     4621 gtttccaatt atattggtga agctaaccag tctgcgtggc tcactgtcac cagacctgtg
     4681 gcaaaagccc tggaagagag gccggcagtg atgacctcgc ccctgtacct ggagatcatc
     4741 atctattgca caggggcctt cctcatctcc tgcatggtgg ggtcggtcat cgtctacaag
     4801 atgaagagtg gtaccaagaa gagtgacttc cacagccaga tggctgtgca caagctggcc
     4861 aagagcatcc ctctgcgcag acaggtaaca gtgtctgctg actccagtgc atccatgaac
     4921 tctggggttc ttctggttcg gccatcacgg ctctcctcca gtgggactcc catgctagca
     4981 ggggtctctg agtatgagct tcccgaagac cctcgctggg agctgcctcg ggacagactg
     5041 gtcttaggca aacccctggg agagggctgc tttgggcagg tggtgttggc agaggctatc
     5101 gggctggaca aggacaaacc caaccgtgtg accaaagtgg ctgtgaagat gttgaagtcg
     5161 gacgcaacag agaaagactt gtcagacctg atctcagaaa tggagatgat gaagatgatc
     5221 gggaagcata agaatatcat caacctgctg ggggcctgca cgcaggatgg tcccttgtat
     5281 gtcatcgtgg agtatgcctc caagggcaac ctgcgggagt acctgcaggc ccggaggccc
     5341 ccagggctgg aatactgcta caaccccagc cacaacccag aggagcagct ctcctccaag
     5401 gacctggtgt cctgcgccta ccaggtggcc cgaggcatgg agtatctggc ctccaagaag
     5461 tgcatacacc gagacctggc agccaggaat gtcctggtga cagaggacaa tgtgatgaag
     5521 atagcagact ttggcctcgc acgggacatt caccacatcg actactataa aaagacaacc
     5581 aacggccaac tgcctgtgaa gtggatggca cccgaggcat tatttgaccg gatctacacc
     5641 caccagagtg atgtgtggtc tttcggggtg ctcctgtggg agatcttcac tctgggcggc
     5701 tccccatacc ccggtgtgcc tgtggaggaa cttttcaagc tgctgaagga gggtcaccgc
     5761 atggacaagc ccagtaactg caccaacgag ctgtacatga tgatgcggga ctgctggcat
     5821 gcagtgccct cacagagacc caccttcaag cagctggtgg aagacctgga ccgcatcgtg
     5881 gccttgacct ccaaccagga gtacctggac ctgtccatgc ccctggacca gtactccccc
     5941 agctttcccg acacccggag ctctacgtgc tcctcagggg aggattccgt cttctctcat
     6001 gagccgctgc ccgaggagcc ctgcctgccc cgacacccag cccagcttgc caatggcgga
     6061 ctcaaacgcc gcgccaaggg tcaagacaat tctgcagata tccagcacag tggcggccgc
     6121 tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc
     6181 gattctacgc gtaccggtca tcatcaccat caccattgag tttaaaccct agagtcgggg
     6241 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     6301 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     6361 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     6421 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     6481 aatcgataag gatccgtttg cgtattgggc gctcttccgc tgatctgcgc agcaccatgg
     6541 cctgaaataa cctctgaaag aggaacttgg ttagctacct tctaaatcgg atccgcggcc
     6601 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     6661 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     6721 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     6781 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     6841 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     6901 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     6961 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     7021 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     7081 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     7141 ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat
     7201 cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga
     7261 gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac
     7321 cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta
     7381 ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac
     7441 ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga
     7501 ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt
     7561 gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg
     7621 cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta
     7681 cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca
     7741 gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct
     7801 gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc
     7861 tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga
     7921 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga
     7981 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     8041 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     8101 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     8161 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     8221 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     8281 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     8341 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     8401 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     8461 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     8521 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc
     8581 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     8641 gctatgtgga tacgctgctt taatgccttt gtatcatgcg ttaactaaac ttgtttattg
     8701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     8761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     8821 attgactcaa atgatgtcaa ttagtctatc agaagctcat ctggtctccc ttccggggga
     8881 caagacatcc ctgtttaata tttaaacagc agtgttccca aactgggttc ttatatccct
     8941 tgctctggtc aaccaggttg cagggtttcc tgtcctcaca ggaacgaagt ccctaaagaa
     9001 acagtggcag ccaggtttag ccccggaatt gactggattc cttttttagg gcccattggt
     9061 atggcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt
     9121 cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg
     9181 ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc
     9241 cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc tgtccctcat
     9301 aaaagttttg ttactttata gaagaaattt tgagtttttg ttttttttaa taaataaata
     9361 aacataaata aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac
     9421 ttaatatcta ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt
     9481 caattttaca catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa
     9541 tctagctgcg tgttctgcag cgtgtcgagc atcttcatct gctccatcac gctgtaaaac
     9601 acatttgcac cgcgagtctg cccgtcctcc acgggttcaa aaacgtgaat gaacgaggcg
     9661 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     9721 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     9781 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     9841 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     9901 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     9961 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10021 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10081 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10141 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10201 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
    10261 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
    10321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
    10381 tgcttcaata atattgaaaa aggaagagt
//
