LOCUS       sequence-231537-       13263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon Destination vector for dox-inducible expression 
            of Gateway cloned elements (inducible mCherry). SpCas9 is 
            constitutively expressed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13263)
  AUTHORS   Kagawa H, Shimamoto R, Kim SI, Oceguera-Yanez F, Yamamoto T, 
            Schroeder T, Woltjen K
  TITLE     OVOL1 Influences the Determination and Expansion of iPSC 
            Reprogramming Intermediates.
  JOURNAL   Stem Cell Reports. 2018 Dec 28. pii: S2213-6711(18)30527-7. doi: 
            10.1016/j.stemcr.2018.12.008.
  PUBMED    30639212
REFERENCE   2  (bases 1 to 13263)
  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: "Stem Cell 
            Reports. 2018 Dec 28. pii: S2213-6711(18)30527-7. doi: 
            10.1016/j.stemcr.2018.12.008."
COMMENT     Sequence Label: PB-TAC-Cas9
FEATURES             Location/Qualifiers
     source          1..13263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   711..773
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1145..1163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1182..1200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1219..1237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1256..1274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1293..1311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1330..1348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1367..1385
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1404..1422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1444..1468
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1548..1672
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1697..1702,1703..1720,1721..1727)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1781..2440
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1847..1866)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2782..3087
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2966..2985
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3128..3252)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(3294..3867,3868..3870,3871..3879)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3461..3478)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3688..3707
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3880..4590
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4025..4043)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4293..4312
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4602..4621)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4667..4722
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4721..4740)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5157..5442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        5444..5721
                     /label=chicken beta-actin promoter
     intron          5722..5950
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5962..5971
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5971..6036
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6043..6063
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6088..10188
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6088..10188
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10189..10236
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(10264..10281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10270..10477
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   10888..10922
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(11072..11090)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11111..11127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11111..11127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11124..11146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11135..11151
                     /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(11159..11165,11166..11183,11184..11189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11204..11225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11342..11359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11513..12101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11593..11612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12272..13063,13064..13132))
                     /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     12895..12914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13133..13237)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tattcacgac agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag
      721 ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta
      781 tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt
      841 acacttacat actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa
      901 aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct
      961 tttacaaaaa taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt
     1021 aattagctta acctatacat aatagaaaca aattatactt attagtcagt cagaaacaac
     1081 tttggcacat atcaatatta tgctctcgtt aattaactcg agggatccaa gctggccgca
     1141 cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga
     1201 aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct
     1261 atcagtgata gagaaggtac gtctagaacg tctccctatc agtgatagag aagtcgacac
     1321 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
     1381 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtaccc cctatataag
     1441 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     1501 ccatagaaga caccgggacc gatccagcct ccgcggtggc ggccatcaca agtttgtaca
     1561 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     1621 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc
     1681 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
     1741 ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     1801 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1861 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1921 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1981 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     2041 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     2101 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     2161 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     2221 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2281 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2341 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     2401 attacaacag tactgcgatg agtggcaggg cggggcgtaa acgcgtggat ccggcttact
     2461 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     2521 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     2581 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     2641 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     2701 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     2761 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     2821 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     2881 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     2941 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     3001 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     3061 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     3121 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     3181 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     3241 gtacaaagtg gtgatctagc taaaacgcgg cctcgaatcg aattccgccc ccccccctct
     3301 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     3361 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     3421 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     3481 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     3541 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3601 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3661 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     3721 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     3781 tttacatgtg tttagtcgag gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3841 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     3901 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3961 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4021 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     4081 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     4141 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     4201 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     4261 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     4321 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     4381 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     4441 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     4501 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     4561 accggcggca tggacgagct gtacaagtaa agcggccgcg atctttttcc ctctgccaaa
     4621 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     4681 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     4741 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     4801 atatgctggc tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc
     4861 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     4921 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     4981 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     5041 aagatccctc gacctgcagg aattcgatta cttaagccta ggatccagat ctgtacattc
     5101 gaatcgatga attcgttaac catggaagct tctgcagcgg ccgcactaga ggtacccgtt
     5161 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5221 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     5281 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     5341 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
     5401 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     5461 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     5521 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg
     5581 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
     5641 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
     5701 aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc
     5761 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     5821 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
     5881 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
     5941 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
     6001 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
     6061 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     6121 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     6181 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     6241 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     6301 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     6361 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     6421 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     6481 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     6541 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     6601 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     6661 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     6721 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     6781 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     6841 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     6901 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     6961 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     7021 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     7081 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     7141 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     7201 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     7261 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     7321 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     7381 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     7441 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     7501 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     7561 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     7621 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     7681 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     7741 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     7801 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     7861 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     7921 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     7981 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     8041 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     8101 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     8161 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     8221 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     8281 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     8341 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     8401 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     8461 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     8521 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     8581 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     8641 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     8701 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     8761 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     8821 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     8881 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     8941 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     9001 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     9061 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     9121 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     9181 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     9241 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     9301 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     9361 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     9421 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     9481 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     9541 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     9601 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     9661 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     9721 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     9781 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     9841 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     9901 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     9961 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
    10021 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
    10081 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
    10141 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
    10201 gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat
    10261 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
    10321 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
    10381 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
    10441 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcactag tctagtctag
    10501 agcgatcgcc cgggtttaaa cgctagccaa caagctcgtc atcgctttgc agaagagcag
    10561 agaggatatg ctcatcgtct aaagaactac ccattttatt atatattagt cacgatatct
    10621 ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa ataacaatat
    10681 aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg tcattttgac
    10741 tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag cacgcctcac
    10801 gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc gcgctattta
    10861 gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact atctttctag
    10921 ggttaaaaaa gatttgcgct ttactcgacc taaactttaa acacgtcata gaatcttcgt
    10981 ttgacaaaaa ccacattgtg gccaagctgt gtgacgcgac gcgcgctaaa gaatggcaaa
    11041 ccaagtcgcg cgaggtaccc agcttttgtt ccctttagtg agggttaatt ccgagcttgg
    11101 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    11161 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    11221 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    11281 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    11341 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    11401 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    11461 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    11521 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    11581 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    11641 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    11701 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    11761 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    11821 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    11881 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    11941 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    12001 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    12061 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    12121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    12181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    12241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    12301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    12361 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    12421 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    12481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    12541 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    12601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    12661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    12721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    12781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    12841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    12901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    12961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    13021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    13081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    13141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    13201 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    13261 ctg
//
