LOCUS       sequence-267625-       11026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for constitutive expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11026)
  AUTHORS   Fischer AD, Veronese-Paniagua DA, Swaminathan S, Kashima H, Rubin 
            DC, Madison BB
  TITLE     The oncogenic function of PLAGL2 is mediated via ASCL2 and IGF2 and 
            a Wnt-independent mechanism in colorectal cancer.
  JOURNAL   Am J Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022.
  PUBMED    37310750
REFERENCE   2  (bases 1 to 11026)
  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: "Am J 
            Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022."
COMMENT     Sequence Label: BII-ChBtW-eSpCas9
FEATURES             Location/Qualifiers
     source          1..11026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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     13..35
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     32..48
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     32..48
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   539..573
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(842..858)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(842..858)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      1791..1800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1797..2192
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             2211..2264
                     /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             2274..2297
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2316..2336
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2361..4898,4899..4901,4902..5363,5364..5366,5367..5534,
                     5535..5537,5538..6461)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6462..6509
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    6536..7124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6589..6609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7007..7018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7204..7338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7228..7247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7282..7301
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   8117..8179
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8309..8326)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8309..8325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8318..8340)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8467..8922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8554..8573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8764..8785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8948..9052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9053..9121,9122..9913)
                     /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(9271..9290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10084..10672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10573..10592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10826..10843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10960..10981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10996..11001,11002..11019,11020..11026)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
       61 aagcgcgccc gccgggtaac tcacggggta tccatgtcca tttctgcggc atccagccag
      121 gatacccgtc ctcgctgacg taatatccca gcgccgcacc gctgtcatta atctgcacac
      181 cggcacggca gttccggctg tcgccggtat tgttcgggtt gctgatgcgc ttcgggctga
      241 ccatccggaa ctgtgtccgg aaaagccgcg acgaactggt atcccaggtg gcctgaacga
      301 acagttcacc gttaaaggcg tgcatggcca caccttcccg aatcatcatg gtaaacgtgc
      361 gttttcgctc aacgtcaatg cagcagcagt catcctcggc aaactctttc catgccgctt
      421 caacctcgcg ggaaaaggca cgggcttctt cctccccgat gcccagatag cgccagcttg
      481 ggcgatgact gagccggaaa aaagacccga cgatatgatc ctgatgcagc tagattaacc
      541 ctagaaagat agtctgcgta aaattgacgc atgcattctt gaaatattgc tctctctttc
      601 taaatagcgc gaatccgtcg ctgtgcattt aggacatctc agtcgccgct tggagctccc
      661 gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt gaactataac gaccgcgtga
      721 gtcaaaatga cgcatgatta tcttttacgt gacttttaag atttaactca tacgataatt
      781 atattgttat ttcatgttct acttacgtga taacttatta tatatatatt ttcttgttat
      841 agatccacta gttctagagg gacagccccc ccccaaagcc cccagggatg taattacgtc
      901 cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc cggcgctccc
      961 cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag gtggcacggg
     1021 atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct ggggggatac
     1081 ggggaaaagg cctccaaggc ctgcaggagt caatgggaaa aacccattgg agccaagtac
     1141 actgactcaa tagggacttt ccattgggtt ttgcccagta cataaggtca atagggggtg
     1201 agtcaacagg aaagtcccat tggagccaag tacattgagt caatagggac tttccaatgg
     1261 gttttgccca gtacataagg tcaatgggag gtaagccaat gggtttttcc cattactgac
     1321 atgtatactg agtcattagg gactttccaa tgggttttgc ccagtacata aggtcaatag
     1381 gggtgaatca acaggaaagt cccattggag ccaagtacac tgagtcaata gggactttcc
     1441 attgggtttt gcccagtaca aaaggtcaat agggggtgag tcaatgggtt tttcccatta
     1501 ttggcacata cataaggtca ataggggtga ctagtggaga agagcatgct tgagggctga
     1561 gtgcccctca gtgggcagag agcacatggc ccacagtccc tgagaagttg gggggagggg
     1621 tgggcaattg aactggtgcc tagagaaggt ggggcttggg taaactggga aagtgatgtg
     1681 gtgtactggc tccacctttt tccccagggt gggggagaac catatataag tgcagtagtc
     1741 tctgtgaaca ttcggtttag tgaacctgca ggtcggccgc cacgaccggt gccaccatgg
     1801 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     1861 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     1921 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     1981 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     2041 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     2101 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     2161 aattgctgcc ctctggttat gtgtgggagg gcgctagcct tggctccggc gagggcaggg
     2221 gaagtcttct aacatgcggg gacgtggagg aaaatcccgg cccactcggc atggactaca
     2281 aggacgacga tgacaagctc gatagcggca tggccccaaa gaagaagcgg aaggtcggta
     2341 tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact
     2401 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     2461 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     2521 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     2581 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     2641 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     2701 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     2761 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     2821 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     2881 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     2941 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     3001 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     3061 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     3121 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc
     3181 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     3241 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     3301 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc
     3361 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     3421 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     3481 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     3541 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc
     3601 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     3661 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     3721 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     3781 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     3841 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     3901 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     3961 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga
     4021 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     4081 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     4141 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     4201 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     4261 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     4321 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     4381 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc
     4441 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     4501 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     4561 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     4621 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     4681 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     4741 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     4801 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     4861 actacgatgt ggaccatatc gtgcctcaga gctttctggc cgacgactcc atcgacaaca
     4921 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     4981 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     5041 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     5101 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     5161 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     5221 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     5281 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     5341 ccgccctgat caaaaagtac cctgcgctgg aaagcgagtt cgtgtacggc gactacaagg
     5401 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     5461 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     5521 gcgagatccg gaaggcgcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     5581 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     5641 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     5701 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     5761 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     5821 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     5881 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     5941 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     6001 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     6061 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     6121 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     6181 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     6241 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     6301 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     6361 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     6421 tgtacgagac acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga
     6481 aaaaggccgg ccaggcaaaa aagaaaaagt aagagctaga gttcgcgtct ggaacaatca
     6541 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6601 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6661 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6721 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6781 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6841 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6901 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     6961 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     7021 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     7081 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgg
     7141 ctaagcactt cgtggccgtc gatcgtttaa agggaggtag tgagtcgaga tatcgatact
     7201 agtgatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     7261 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     7321 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     7381 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     7441 gatcctctag agggacagcc cccccccaaa gcccccaggg atgtaattac gtccctcccc
     7501 cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct ccccccgcat
     7561 ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac gggatcgctt
     7621 tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga tacggggaaa
     7681 aagccatacc aatggagctc acggggacag ccccccccca aagcccccag ggatgtaatt
     7741 acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg
     7801 ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc
     7861 acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg
     7921 gatacgggga aaaagaattc gataaaagtt ttgttacttt atagaagaaa ttttgagttt
     7981 ttgttttttt taataaataa ataaacataa ataaattgtt tgttgaattt attattagta
     8041 tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa acctcgatat
     8101 acagaccgat aaaacacatg cgtcaatttt acacatgatt atctttaacg tacgtcacaa
     8161 tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg agcatcttca
     8221 tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc tccacgggtt
     8281 caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     8341 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     8401 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     8461 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     8521 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8581 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8641 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     8701 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     8761 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     8821 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8881 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     8941 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     9001 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     9061 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     9121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     9181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     9241 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     9301 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     9361 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     9421 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     9481 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     9541 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     9601 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     9661 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     9721 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     9781 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9841 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9901 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9961 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
    10021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    10081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
    10141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
    10201 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
    10261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
    10321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
    10381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
    10441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
    10501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    10561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    10621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    10681 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    10741 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    10801 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
    10861 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
    10921 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
    10981 ttaggcaccc caggctttac actttatgct tccggctcgt atgttg
//
