LOCUS       sequence-153664-       12468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-One piggyBac transposon Destination vector for dox-inducible 
            expression of Gateway cloned elements (inducible GFP and 
            constitutive rtTA and neomycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12468)
  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 12468)
  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-TAG-ERN
FEATURES             Location/Qualifiers
     source          1..12468
                     /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"
     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..3868,3869..3871,3872..3880)
                     /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             join(3881..3883,3884..3886,3887..4600)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3881..3883,3884..3886,3887..4597)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3926..3947)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4187..4206)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4534..4555
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4612..4631)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4677..4732
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4731..4750)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5173..5552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5667..6972
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    6009..6940
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(6990..7607,7608..7613,7614..7649,7650..7652,7653..7688,
                     7689..7691,7692..7727,7728..7736)
                     /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    7763..8207
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        8240..8286
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        8278..8296
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8306..9100
                     /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             8306..9100
                     /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(8360..8379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8970..8989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9147..9719
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   10093..10127
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(10277..10295)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10316..10332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10316..10332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10329..10351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10340..10356
                     /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(10364..10370,10371..10388,10389..10394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10409..10430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10547..10564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10718..11306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10798..10817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11477..12268,12269..12337))
                     /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     12100..12119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12338..12442)
                     /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 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     3841 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     3901 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3961 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     4021 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     4081 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4141 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4201 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4261 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4321 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     4381 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4441 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     4501 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4561 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg atctttttcc
     4621 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     4681 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     4741 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     4801 aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt catcagtata
     4861 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     4921 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     4981 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     5041 ttctcttatg aagatccctc gacctgcagg aattcgatta cttaagccta ggatccagat
     5101 ctgtacattc gaatcgatga attcgttaac catggaagct tctgcagcgg ccgcactagc
     5161 ggtggcggcc tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     5221 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     5281 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     5341 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     5401 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     5461 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5521 acatctacgt attagtcatc gctattacca tgatgatgcg gttttggcag tacatcaatg
     5581 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     5641 ggagtttgtt ttgactagtg gagccgagag taattcatac aaaaggaggg atcgccttcg
     5701 caaggggaga gcccagggac cgtccctaaa ttctcacaga cccaaatccc tgtagccgcc
     5761 ccacgacagc gcgaggagca tgcgcccagg gctgagcgcg ggtagatcag agcacacaag
     5821 ctcacagtcc ccggcggtgg ggggaggggc gcgctgagcg ggggccaggg agctggcgcg
     5881 gggcaaactg ggaaagtggt gtcgtgtgct ggctccgccc tcttcccgag ggtgggggag
     5941 aacggtatat aagtgcggta gtcgccttgg acgttctttt tcgcaacggg tttgccgtca
     6001 gaacgcaggt gagtggcggg tgtggcttcc gcgggccccg gagctggagc cctgctctga
     6061 gcgggccggg ctgatatgcg agtgtcgtcc gcagggttta gctgtgagca ttcccacttc
     6121 gagtggcggg cggtgcgggg gtgagagtgc gaggcctagc ggcaaccccg tagcctcgcc
     6181 tcgtgtccgg cttgaggcct agcgtggtgt ccgccgccgc gtgccactcc ggccgcacta
     6241 tgcgtttttt gtccttgctg ccctcgattg ccttccagca gcatgggcta acaaagggag
     6301 ggtgtggggc tcactcttaa ggagcccatg aagcttacgt tggataggaa tggaagggca
     6361 ggaggggcga ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg
     6421 cgaggcctgt ggctttccga agcaatcggg cgtgagttta gcctacctgg gccatgtggc
     6481 cctagcactg ggcacggtct ggcctggcgg tgccgcgttc ccttgcctcc caacaagggt
     6541 gaggccgtcc cgcccggcac cagttgcttg cgcggaaaga tggccgctcc cggggccctg
     6601 ttgcaaggag ctcaaaatgg aggacgcggc agcccggtgg agcgggcggg tgagtcaccc
     6661 acacaaagga agagggcctt gcccctcgcc ggccgctgct tcctgtgacc ccgtggtcta
     6721 tcggccgcat agtcacctcg ggcttctctt gagcaccgct cgtcgcggcg gggggagggg
     6781 atctaatggc gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg
     6841 cgctggaagt cattcttgga atttgcccct ttgagtttgg agcgaggcta attctcaagc
     6901 ctcttagcgg ttcaaaggta ttttctaaac ccgtttccag gtgttgtgaa agccaccgct
     6961 aattcaaagc aatccggagt aaattcacca tgtctagact ggacaagagc aaagtcataa
     7021 acggcgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     7081 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     7141 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     7201 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc
     7261 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     7321 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     7381 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     7441 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     7501 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     7561 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccggccgacg
     7621 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     7681 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa
     7741 gtaaggatcc cgtacgccta ggagcaggtt tccccaatga cacaaaacgt gcaacttgaa
     7801 actccgcctg gtctttccag gtctagaggg gtaacacttt gtactgcgtt tggctccacg
     7861 ctcgatccac tggcgagtgt tagtaacagc actgttgctt cgtagcggag catgacggcc
     7921 gtgggaactc ctccttggta acaaggaccc acggggccaa aagccacgcc cacacgggcc
     7981 cgtcatgtgt gcaaccccag cacggcgact ttactgcgaa acccacttta aagtgacatt
     8041 gaaactggta cccacacact ggtgacaggc taaggatgcc cttcaggtac cccgaggtaa
     8101 cacgcgacac tcgggatctg agaaggggac tggggcttct ataaaagcgc tcggtttaaa
     8161 aagcttctat gcctgaatag gtgaccggag gtcggcacct ttcctttgca attactgacc
     8221 ctatgaatac actgactgtt tgacaattaa tcatcggcat agtatatcgg catagtataa
     8281 tacgactcac tataggaggg ccaccatgat tgaacaagat ggattgcacg caggttctcc
     8341 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     8401 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     8461 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     8521 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     8581 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     8641 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     8701 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     8761 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     8821 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacacatg gcgatgcctg
     8881 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     8941 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     9001 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     9061 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     9121 atgaccgacc aagcgaattc gctaggatta tccctaatac ctgccacccc actcttaatc
     9181 agtggtggaa gaacggtctc agaactgttt gtttcaattg gccatttaag tttagtagta
     9241 aaagactggt taatgataac aatgcatcgt aaaaccttca gaaggaaagg agaatgtttt
     9301 gtggaccact ttggttttct tttttgcgtg tggcagtttt aagttattag tttttaaaat
     9361 cagtactttt taatggaaac aacttgacca aaaatttgtc acagaatttt gagacccatt
     9421 aaaaaagtta aatgagaaac ctgtgtgttc ctttggtcaa caccgagaca tttaggtgaa
     9481 agacatctaa ttctggtttt acgaatctgg aaacttcttg aaaatgtaat tcttgagtta
     9541 acacttctgg gtggagaata gggttgtttt ccccccacat aattggaagg ggaaggaata
     9601 tcatttaaag ctatgggagg gttgctttga ttacaacact ggagagaaat gcagcatgtt
     9661 gctgattgcc tgtcactaaa acaggccaaa aactgagtcc ttgggttgca tagaaagctg
     9721 ccaaacgcta gccaacaagc tcgtcatcgc tttgcagaag agcagagagg atatgctcat
     9781 cgtctaaaga actacccatt ttattatata ttagtcacga tatctataac aagaaaatat
     9841 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     9901 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     9961 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
    10021 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
    10081 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta aaaaagattt
    10141 gcgctttact cgacctaaac tttaaacacg tcatagaatc ttcgtttgac aaaaaccaca
    10201 ttgtggccaa gctgtgtgac gcgacgcgcg ctaaagaatg gcaaaccaag tcgcgcgagg
    10261 tacccagctt ttgttccctt tagtgagggt taattccgag cttggcgtaa tcatggtcat
    10321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    10381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    10441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    10501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    10561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    10621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    10681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    10741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    10801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    10861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    10921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    10981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    11041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    11101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
    11161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    11221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    11281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
    11341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
    11401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
    11461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
    11521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
    11581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    11641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    11701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    11761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    11821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    11881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    11941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    12001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    12061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    12121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    12181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    12241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    12301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    12361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    12421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
