LOCUS       sequence-256350-       11025 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 11025)
  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 11025)
  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..11025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(308..324)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      1257..1266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1263..1658
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1677..1730
                     /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             1740..1763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1782..1802
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1827..4364,4365..4367,4368..4829,4830..4832,4833..5000,
                     5001..5003,5004..5927)
                     /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             5928..5975
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    6002..6590
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6055..6075)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6473..6484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6670..6804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6694..6713)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6748..6767
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7582..7644
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7774..7791)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7774..7790)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7783..7805)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7932..8387
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8019..8038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8229..8250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8413..8517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8518..8586,8587..9378)
                     /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(8736..8755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9549..10137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10038..10057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10291..10308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10425..10446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10461..10466,10467..10484,10485..10491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10499..10515
                     /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     10504..10526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10523..10539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10523..10539
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat
      361 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
      421 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
      481 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
      541 ggatacgggg aaaaggcctc caaggcctgc aggagtcaat gggaaaaacc cattggagcc
      601 aagtacactg actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag
      661 ggggtgagtc aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc
      721 caatgggttt tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt
      781 actgacatgt atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt
      841 caataggggt gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga
      901 ctttccattg ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc
      961 ccattattgg cacatacata aggtcaatag gggtgactag tggagaagag catgcttgag
     1021 ggctgagtgc ccctcagtgg gcagagagca catggcccac agtccctgag aagttggggg
     1081 gaggggtggg caattgaact ggtgcctaga gaaggtgggg cttgggtaaa ctgggaaagt
     1141 gatgtggtgt actggctcca cctttttccc cagggtgggg gagaaccata tataagtgca
     1201 gtagtctctg tgaacattcg gtttagtgaa cctgcaggtc ggccgccacg accggtgcca
     1261 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     1321 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     1381 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     1441 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     1501 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     1561 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     1621 ttcgtgaatt gctgccctct ggttatgtgt gggagggcgc tagccttggc tccggcgagg
     1681 gcaggggaag tcttctaaca tgcggggacg tggaggaaaa tcccggccca ctcggcatgg
     1741 actacaagga cgacgatgac aagctcgata gcggcatggc cccaaagaag aagcggaagg
     1801 tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca
     1861 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     1921 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     1981 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     2041 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     2101 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     2161 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     2221 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     2281 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     2341 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     2401 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     2461 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     2521 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     2581 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     2641 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     2701 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     2761 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     2821 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     2881 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     2941 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     3001 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     3061 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     3121 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     3181 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     3241 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     3301 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     3361 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     3421 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     3481 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     3541 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     3601 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     3661 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     3721 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     3781 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     3841 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     3901 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     3961 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     4021 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     4081 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     4141 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     4201 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     4261 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     4321 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctggccgac gactccatcg
     4381 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     4441 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     4501 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     4561 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     4621 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     4681 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     4741 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     4801 tgggaaccgc cctgatcaaa aagtaccctg cgctggaaag cgagttcgtg tacggcgact
     4861 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     4921 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     4981 ccaacggcga gatccggaag gcgcctctga tcgagacaaa cggcgaaacc ggggagatcg
     5041 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     5101 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     5161 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     5221 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     5281 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     5341 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     5401 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     5461 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     5521 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     5581 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     5641 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     5701 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     5761 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     5821 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     5881 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg
     5941 ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaaga gctagagttc gcgtctggaa
     6001 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6061 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6121 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6181 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6241 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6301 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6361 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6421 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6481 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6541 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     6601 ttctggctaa gcacttcgtg gccgtcgatc gtttaaaggg aggtagtgag tcgagatatc
     6661 gatactagtg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     6721 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     6781 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     6841 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     6901 gattatgatc ctctagaggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
     6961 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
     7021 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
     7081 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
     7141 gggaaaagcc ataccaatgg agctcacggg gacagccccc ccccaaagcc cccagggatg
     7201 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
     7261 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
     7321 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
     7381 ggggggatac ggggaaaaag aattcgataa aagttttgtt actttataga agaaattttg
     7441 agtttttgtt ttttttaata aataaataaa cataaataaa ttgtttgttg aatttattat
     7501 tagtatgtaa gtgtaaatat aataaaactt aatatctatt caaattaata aataaacctc
     7561 gatatacaga ccgataaaac acatgcgtca attttacaca tgattatctt taacgtacgt
     7621 cacaatatga ttatctttct agggttaatc tagctgcgtg ttctgcagcg tgtcgagcat
     7681 cttcatctgc tccatcacgc tgtaaaacac atttgcaccg cgagtctgcc cgtcctccac
     7741 gggttcaaaa acgtgaatga acgaggcgcg ctcactggcc gtcgttttac aacgtcgtga
     7801 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     7861 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     7921 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     7981 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     8041 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     8101 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     8161 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     8221 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     8281 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     8341 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     8401 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     8461 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     8521 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     8581 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     8641 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     8701 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     8761 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     8821 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8881 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8941 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     9001 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     9061 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     9121 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     9181 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     9241 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     9301 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     9361 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     9421 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     9481 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     9541 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     9601 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     9661 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     9721 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9781 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9841 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9901 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9961 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    10021 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    10081 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    10141 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    10201 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    10261 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
    10321 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
    10381 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
    10441 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
    10501 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgcc
    10561 cgccgggtaa ctcacggggt atccatgtcc atttctgcgg catccagcca ggatacccgt
    10621 cctcgctgac gtaatatccc agcgccgcac cgctgtcatt aatctgcaca ccggcacggc
    10681 agttccggct gtcgccggta ttgttcgggt tgctgatgcg cttcgggctg accatccgga
    10741 actgtgtccg gaaaagccgc gacgaactgg tatcccaggt ggcctgaacg aacagttcac
    10801 cgttaaaggc gtgcatggcc acaccttccc gaatcatcat ggtaaacgtg cgttttcgct
    10861 caacgtcaat gcagcagcag tcatcctcgg caaactcttt ccatgccgct tcaacctcgc
    10921 gggaaaaggc acgggcttct tcctccccga tgcccagata gcgccagctt gggcgatgac
    10981 tgagccggaa aaaagacccg acgatatgat cctgatgcag ctaga
//
