LOCUS       sequence-477521-       12453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-One piggyBac transposon Destination vector for dox-inducible 
            expression of Gateway cloned elements (inducible mCherry and 
            constitutive rtTA and neomycin resistance) aka. KW111.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12453)
  AUTHORS   Kim SI, Oceguera-Yanez F, Sakurai C, Nakagawa M, Yamanaka S, Woltjen
            K
  TITLE     Inducible Transgene Expression in Human iPS Cells Using Versatile 
            All-in-One piggyBac Transposons.
  JOURNAL   Methods Mol Biol. 2016;1357:111-31. doi: 10.1007/7651_2015_251.
  PUBMED    26025620
REFERENCE   2  (bases 1 to 12453)
  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: "Methods Mol
            Biol."; date: "2016"; volume: "1357:111-31. doi"; pages: " 
            10.1007/7651_2015_251"
COMMENT     Sequence Label: PB-TAC-ERN
FEATURES             Location/Qualifiers
     source          1..12453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..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     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   708..770
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1142..1160
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1179..1197
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1216..1234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1253..1271
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1290..1308
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1327..1345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1364..1382
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1401..1419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1441..1465
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1545..1669
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1694..1699,1700..1717,1718..1724)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1778..2437
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1844..1863)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2779..3084
                     /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     2963..2982
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3125..3249)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(3291..3865,3866..3868,3869..3877)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3458..3475)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3685..3704
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3878..4588
                     /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(4023..4041)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4291..4310
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4600..4619)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4665..4720
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4719..4738)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5163..5542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5657..6962
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    5999..6930
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(6979..7596,7597..7602,7603..7638,7639..7641,7642..7677,
                     7678..7680,7681..7716,7717..7725)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    7752..8196
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        8229..8275
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        8267..8285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8295..9089
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             8295..9089
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8349..8368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8959..8978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9136..9708
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   10082..10116
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(10265..10283)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10304..10320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10304..10320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10317..10339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10328..10344
                     /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(10352..10358,10359..10376,10377..10382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10397..10418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10535..10552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10706..11294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10786..10805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11465..12256,12257..12325))
                     /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     12088..12107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12326..12430)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata 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 gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctcggtat
      661 tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc tagaaagata
      721 atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg
      781 gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca
      841 cttacatact aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa
      901 aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat tctctctttt
      961 acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa aataagtaat
     1021 tagcttaacc tatacataat agaaacaaat tatacttatt agtcagtcag aaacaacttt
     1081 ggcacatatc aatattatgc tctcgttaat taactcgagg gatccaagct ggccgcacgt
     1141 ctccctatca gtgatagaga agtcgacacg tctcgagctc cctatcagtg atagagaagg
     1201 tacgtctaga acgtctccct atcagtgata gagaagtcga cacgtctcga gctccctatc
     1261 agtgatagag aaggtacgtc tagaacgtct ccctatcagt gatagagaag tcgacacgtc
     1321 tcgagctccc tatcagtgat agagaaggta cgtctagaac gtctccctat cagtgataga
     1381 gaagtcgaca cgtctcgagc tccctatcag tgatagagaa ggtaccccct atataagcag
     1441 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     1501 tagaagacac cgggaccgat ccagcctccg cggtggcggc catcacaagt ttgtacaaaa
     1561 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     1621 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt
     1681 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
     1741 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     1801 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     1861 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     1921 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     1981 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     2041 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     2101 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     2161 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     2221 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     2281 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     2341 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     2401 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa
     2461 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     2521 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     2581 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     2641 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     2701 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     2761 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     2821 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     2881 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     2941 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     3001 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     3061 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     3121 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     3181 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     3241 caaagtggtg atctagctaa aacgcggcct cgaatcgaat tccgcccccc cccctctccc
     3301 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     3361 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     3421 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     3481 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     3541 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     3601 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     3661 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     3721 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     3781 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     3841 ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagga
     3901 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     3961 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     4021 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     4081 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     4141 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     4201 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     4261 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     4321 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     4381 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     4441 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     4501 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     4561 cggcggcatg gacgagctgt acaagtaaag cggccgcgat ctttttccct ctgccaaaaa
     4621 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     4681 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     4741 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     4801 tatgctggct gccatgaaca aaggttggct 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 gagatccctc gacctgcagg aattcgatta cttaagccta ggatccagat ctgtacattc
     5101 gaatcgatga attcgttaac catggaagct tctgcagcgg ccgcactagc ggtggcggcc
     5161 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     5221 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5281 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     5341 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     5401 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     5461 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     5521 attagtcatc gctattacca tgatgatgcg gttttggcag tacatcaatg ggcgtggata
     5581 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     5641 ttgactagtg gagccgagag taattcatac aaaaggaggg atcgccttcg caaggggaga
     5701 gcccagggac cgtccctaaa ttctcacaga cccaaatccc tgtagccgcc ccacgacagc
     5761 gcgaggagca tgcgcccagg gctgagcgcg ggtagatcag agcacacaag ctcacagtcc
     5821 ccggcggtgg ggggaggggc gcgctgagcg ggggccaggg agctggcgcg gggcaaactg
     5881 ggaaagtggt gtcgtgtgct ggctccgccc tcttcccgag ggtgggggag aacggtatat
     5941 aagtgcggta gtcgccttgg acgttctttt tcgcaacggg tttgccgtca gaacgcaggt
     6001 gagtggcggg tgtggcttcc gcgggccccg gagctggagc cctgctctga gcgggccggg
     6061 ctgatatgcg agtgtcgtcc gcagggttta gctgtgagca ttcccacttc gagtggcggg
     6121 cggtgcgggg gtgagagtgc gaggcctagc ggcaaccccg tagcctcgcc tcgtgtccgg
     6181 cttgaggcct agcgtggtgt ccgccgccgc gtgccactcc ggccgcacta tgcgtttttt
     6241 gtccttgctg ccctcgattg ccttccagca gcatgggcta acaaagggag ggtgtggggc
     6301 tcactcttaa ggagcccatg aagcttacgt tggataggaa tggaagggca ggaggggcga
     6361 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgt
     6421 ggctttccga agcaatcggg cgtgagttta gcctacctgg gccatgtggc cctagcactg
     6481 ggcacggtct ggcctggcgg tgccgcgttc ccttgcctcc caacaagggt gaggccgtcc
     6541 cgcccggcac cagttgcttg cgcggaaaga tggccgctcc cggggccctg ttgcaaggag
     6601 ctcaaaatgg aggacgcggc agcccggtgg agcgggcggg tgagtcaccc acacaaagga
     6661 agagggcctt gcccctcgcc ggccgctgct tcctgtgacc ccgtggtcta tcggccgcat
     6721 agtcacctcg ggcttctctt gagcaccgct cgtcgcggcg gggggagggg atctaatggc
     6781 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     6841 cattcttgga atttgcccct ttgagtttgg agcgaggcta attctcaagc ctcttagcgg
     6901 ttcaaaggta ttttctaaac ccgtttccag gtgttgtgaa agccaccgct aattcaaagc
     6961 aatccggagt aattcaccat gtctagactg gacaagagca aagtcataaa cggcgctctg
     7021 gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     7081 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     7141 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     7201 tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct ctcacatcgc
     7261 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     7321 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     7381 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     7441 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     7501 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     7561 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cggccgacgc ccttgacgat
     7621 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     7681 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag taaggatccc
     7741 gtacgcctag gagcaggttt ccccaatgac acaaaacgtg caacttgaaa ctccgcctgg
     7801 tctttccagg tctagagggg taacactttg tactgcgttt ggctccacgc tcgatccact
     7861 ggcgagtgtt agtaacagca ctgttgcttc gtagcggagc atgacggccg tgggaactcc
     7921 tccttggtaa caaggaccca cggggccaaa agccacgccc acacgggccc gtcatgtgtg
     7981 caaccccagc acggcgactt tactgcgaaa cccactttaa agtgacattg aaactggtac
     8041 ccacacactg gtgacaggct aaggatgccc ttcaggtacc ccgaggtaac acgcgacact
     8101 cgggatctga gaaggggact ggggcttcta taaaagcgct cggtttaaaa agcttctatg
     8161 cctgaatagg tgaccggagg tcggcacctt tcctttgcaa ttactgaccc tatgaataca
     8221 ctgactgttt gacaattaat catcggcata gtatatcggc atagtataat acgactcact
     8281 ataggagggc caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     8341 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     8401 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     8461 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     8521 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     8581 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     8641 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     8701 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     8761 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     8821 cgagcatgcc cgacggcgag gatctcgtcg tgacacatgg cgatgcctgc ttgccgaata
     8881 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     8941 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     9001 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     9061 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     9121 agcgaattcg ctaggattat ccctaatacc tgccacccca ctcttaatca gtggtggaag
     9181 aacggtctca gaactgtttg tttcaattgg ccatttaagt ttagtagtaa aagactggtt
     9241 aatgataaca atgcatcgta aaaccttcag aaggaaagga gaatgttttg tggaccactt
     9301 tggttttctt ttttgcgtgt ggcagtttta agttattagt ttttaaaatc agtacttttt
     9361 aatggaaaca acttgaccaa aaatttgtca cagaattttg agacccatta aaaaagttaa
     9421 atgagaaacc tgtgtgttcc tttggtcaac accgagacat ttaggtgaaa gacatctaat
     9481 tctggtttta cgaatctgga aacttcttga aaatgtaatt cttgagttaa cacttctggg
     9541 tggagaatag ggttgttttc cccccacata attggaaggg gaaggaatat catttaaagc
     9601 tatgggaggg ttgctttgat tacaacactg gagagaaatg cagcatgttg ctgattgcct
     9661 gtcactaaaa caggccaaaa actgagtcct tgggttgcat agaaagctgc caaacgctag
     9721 ccaacaagct cgtcatcgct ttgcagaaga gcagagagga tatgctcatc gtctaaagaa
     9781 ctacccattt tattatatat tagtcacgat atctataaca agaaaatata tatataataa
     9841 gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa
     9901 agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc
     9961 agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg
    10021 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
    10081 gcatgcgtca attttacgca gactatcttt ctagggttaa aaaagatttg cgctttactc
    10141 gacctaaact ttaaacacgt catagaatct tcgtttgaca aaaaccacat tgtggccaag
    10201 ctgtgtgacg cgacgcgcgc taaagaatgg caaaccaagt cgcgcgaggt accagctttt
    10261 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
    10321 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
    10381 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
    10441 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
    10501 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
    10561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
    10621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
    10681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    10741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    10801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    10861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    10921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    10981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    11041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    11101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
    11161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    11221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    11281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    11341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
    11401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
    11461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
    11521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
    11581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
    11641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    11701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    11761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    11821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    11881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    11941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    12001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    12061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    12121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    12181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    12241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    12301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    12361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    12421 gggttccgcg cacatttccc cgaaaagtgc cac
//
