LOCUS       sequence-274605-       12733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac expression vector with TET-ON system and Blasticidin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12733)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 12733)
  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"
COMMENT     Sequence Label: PB-TET-BH-lox66FRT
FEATURES             Location/Qualifiers
     source          1..12733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   706..768
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     1177..1193
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1204..1703
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1239..1262)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1636..1655
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     polyA_signal    2235..2369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2272..2291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2326..2345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2677..2693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2701..2717
                     /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(2725..2731,2732..2749,2750..2755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2770..2791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    2874..2892
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2911..2929
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2948..2966
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2985..3003
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3022..3040
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3059..3077
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3096..3114
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3133..3151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3173..3197
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3277..3401
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3426..3431,3432..3449,3450..3456)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3510..4169
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3576..3595)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4511..4816
                     /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     4695..4714
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4857..4981)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    5033..5606
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5056..5608
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5200..5217)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5427..5446
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5647..8688
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQ"
     CDS             8713..9516
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8776..8795)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9386..9405
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(9548..9565)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9554..9778
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   10217..10251
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    complement(10278..10311)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    10344..10391
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(10543..10561)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10582..10598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10606..10622
                     /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(10630..10636,10637..10654,10655..10660))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10675..10696
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10813..10830)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10984..11572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11064..11083)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11743..12534,12535..12603))
                     /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     12366..12385
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12604..12708)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttactgg cgaaaggggg atgtgctgca aggcgattaa
     1141 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattct
     1201 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     1261 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     1321 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     1381 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     1441 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     1501 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     1561 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     1621 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     1681 cttcctcatc tccgggcctt tcgaccgatc atcagatcta ccatgccttt gtctcaagaa
     1741 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     1801 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     1861 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     1921 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     1981 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     2041 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     2101 gtgtgggagg gttaactaga taagtaatga tcataatcag ccatatcaca tctgtagagg
     2161 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2221 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2281 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2341 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaaatttaa ctctagagag
     2401 ctcacgggga cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta
     2461 gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg
     2521 agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc
     2581 tgaacgcttc tcgctgctct ttgagcctgc agacacctgg gggatacggg gaaaaagctt
     2641 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2821 gtcgggaaac ctgtcgtgcc agtaactcga gggatccaag ctggccgcac gtctccctat
     2881 cagtgataga gaagtcgaca cgtctcgagc tccctatcag tgatagagaa ggtacgtcta
     2941 gaacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta tcagtgatag
     3001 agaaggtacg tctagaacgt ctccctatca gtgatagaga agtcgacacg tctcgagctc
     3061 cctatcagtg atagagaagg tacgtctaga acgtctccct atcagtgata gagaagtcga
     3121 cacgtctcga gctccctatc agtgatagag aaggtacccc ctatataagc agagctcgtt
     3181 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
     3241 accgggaccg atccagcctc cgcggtggcg gccatcacaa gtttgtacaa aaaagctgaa
     3301 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
     3361 actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc
     3421 caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt
     3481 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     3541 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     3601 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     3661 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     3721 cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     3781 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     3841 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     3901 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
     3961 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     4021 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     4081 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     4141 actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta aaagccagat
     4201 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
     4261 cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag
     4321 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
     4381 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
     4441 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
     4501 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     4561 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
     4621 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     4681 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     4741 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     4801 gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata
     4861 gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat
     4921 ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg
     4981 tgatggccgc tctagctagt cgataagcta attccgcccc cccccccccc cccccctctc
     5041 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     5101 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     5161 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     5221 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     5281 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     5341 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     5401 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     5461 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     5521 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     5581 ttttcctttg aaaaacacga tgataatatg gccatgagct tgatgatctg tgacatggcg
     5641 gatcccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     5701 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     5761 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
     5821 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
     5881 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc
     5941 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
     6001 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
     6061 aactcggcgt ttcatctgtg gtgcaacggg cgctgggtcg gttacggcca ggacagtcgt
     6121 ttgccgtctg aatttgacct gagcgcattt ttacgcgccg gagaaaaccg cctcgcggtg
     6181 atggtgctgc gctggagtga cggcagttat ctggaagatc aggatatgtg gcggatgagc
     6241 ggcattttcc gtgacgtctc gttgctgcat aaaccgacta cacaaatcag cgatttccat
     6301 gttgccactc gctttaatga tgatttcagc cgcgctgtac tggaggctga agttcagatg
     6361 tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg tgaaacgcag
     6421 gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg atgagcgtgg tggttatgcc
     6481 gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc cgaaatcccg
     6541 aatctctatc gtgcggtggt tgaactgcac accgccgacg gcacgctgat tgaagcagaa
     6601 gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct gctgaacggc
     6661 aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca tggtcaggtc
     6721 atggatgagc agacgatggt gcaggatatc ctgctgatga agcagaacaa ctttaacgcc
     6781 gtgcgctgtt cgcattatcc gaaccatccg ctgtggtaca cgctgtgcga ccgctacggc
     6841 ctgtatgtgg tggatgaagc caatattgaa acccacggca tggtgccaat gaatcgtctg
     6901 accgatgatc cgcgctggct accggcgatg agcgaacgcg taacgcgaat ggtgcagcgc
     6961 gatcgtaatc acccgagtgt gatcatctgg tcgctgggga atgaatcagg ccacggcgct
     7021 aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc cttcccgccc ggtgcagtat
     7081 gaaggcggcg gagccgacac cacggccacc gatattattt gcccgatgta cgcgcgcgtg
     7141 gatgaagacc agcccttccc ggctgtgccg aaatggtcca tcaaaaaatg gctttcgcta
     7201 cctggagaga cgcgcccgct gatcctttgc gaatacgccc acgcgatggg taacagtctt
     7261 ggcggtttcg ctaaatactg gcaggcgttt cgtcagtatc cccgtttaca gggcggcttc
     7321 gtctgggact gggtggatca gtcgctgatt aaatatgatg aaaacggcaa cccgtggtcg
     7381 gcttacggcg gtgattttgg cgatacgccg aacgatcgcc agttctgtat gaacggtctg
     7441 gtctttgccg accgcacgcc gcatccagcg ctgacggaag caaaacacca gcagcagttt
     7501 ttccagttcc gtttatccgg gcaaaccatc gaagtgacca gcgaatacct gttccgtcat
     7561 agcgataacg agctcctgca ctggatggtg gcgctggatg gtaagccgct ggcaagcggt
     7621 gaagtgcctc tggatgtcgc tccacaaggt aaacagttga ttgaactgcc tgaactaccg
     7681 cagccggaga gcgccgggca actctggctc acagtacgcg tagtgcaacc gaacgcgacc
     7741 gcatggtcag aagccgggca catcagcgcc tggcagcagt ggcgtctggc ggaaaacctc
     7801 agtgtgacgc tccccgccgc gtcccacgcc atcccgcatc tgaccaccag cgaaatggat
     7861 ttttgcatcg agctgggtaa taagcgttgg caatttaacc gccagtcagg ctttctttca
     7921 cagatgtgga ttggcgataa aaaacaactg ctgacgccgc tgcgcgatca gttcacccgt
     7981 gcaccgctgg ataacgacat tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg
     8041 gtcgaacgct ggaaggcggc gggccattac caggccgaag cagcgttgtt gcagtgcacg
     8101 gcagatacac ttgctgatgc ggtgctgatt acgaccgctc acgcgtggca gcatcagggg
     8161 aaaaccttat ttatcagccg gaaaacctac cggattgatg gtagtggtca aatggcgatt
     8221 accgttgatg ttgaagtggc gagcgataca ccgcatccgg cgcggattgg cctgaactgc
     8281 cagctggcgc aggtagcaga gcgggtaaac tggctcggat tagggccgca agaaaactat
     8341 cccgaccgcc ttactgccgc ctgttttgac cgctgggatc tgccattgtc agacatgtat
     8401 accccgtacg tcttcccgag cgaaaacggt ctgcgctgcg ggacgcgcga attgaattat
     8461 ggcccacacc agtggcgcgg cgacttccag ttcaacatca gccgctacag tcaacagcaa
     8521 ctgatggaaa ccagccatcg ccatctgctg cacgcggaag aaggcacatg gctgaatatc
     8581 gacggtttcc atatggggat tggtggcgac gactcctgga gcccgtcagt atcggcggaa
     8641 ttccagctga gcgccggtcg ctaccattac cagttggtct ggtgtcaggg gatcccccgg
     8701 gctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc
     8761 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     8821 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     8881 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     8941 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     9001 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     9061 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     9121 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     9181 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     9241 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     9301 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     9361 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     9421 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     9481 atcgccttct atcgccttct tgacgagttc ttctgagggg atcaattctc tagagctcgc
     9541 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     9601 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     9661 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     9721 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     9781 tctgaggcgg aaagaaccag ctggggctcg atcctctaga gtcgagggct gcaggaattc
     9841 gatatcaagc tctagccaac aagctcgtca tcgctttgca gaagagcaga gaggatatgc
     9901 tcatcgtcta aagaactacc cattttatta tatattagtc acgatatcta taacaagaaa
     9961 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
    10021 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
    10081 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
    10141 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
    10201 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaaatttg
    10261 ggcggtcgag ggacctaata acttcgtata gcatacatta tacgaacggt aattaagggt
    10321 tccggatccc catcaagcta attgaagttc ctattccgaa gttcctattc tctagaaagt
    10381 ataggaactt ccagctggaa atgatttgcg ctttactcga cctaaacttt aaacacgtca
    10441 tagaatcttc gtttgacaaa aaccacattg tggccaagct gtgtgacgcg acgcgcgcta
    10501 aagaatggca aaccaagtcg cgcgaggtac cagcttttgt tccctttagt gagggttaat
    10561 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    10621 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    10681 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    10741 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    10801 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    10861 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    10921 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    10981 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    11041 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    11101 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    11161 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    11221 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    11281 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    11341 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    11401 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
    11461 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    11521 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    11581 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    11641 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    11701 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    11761 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    11821 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    11881 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    11941 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    12001 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    12061 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    12121 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    12181 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    12241 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    12301 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    12361 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    12421 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    12481 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    12541 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    12601 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    12661 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    12721 aaaagtgcca cct
//
