LOCUS       sequence-128108-       18186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rosa26 targeting vector expressing Cre-dependent tTR-KRAB-rtTA3-Luc 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18186)
  AUTHORS   Akama-Garren EH, Joshi NS, Tammela T, Chang GP, Wagner BL, Lee DY, 
            Rideout Iii WM, Papagiannakopoulos T, Xue W, Jacks T
  TITLE     A Modular Assembly Platform for Rapid Generation of DNA Constructs.
  JOURNAL   Sci Rep. 2016 Feb 18;6:16836. doi: 10.1038/srep16836.
  PUBMED    26887506
REFERENCE   2  (bases 1 to 18186)
  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: "Sci Rep."; 
            date: "2016-02-18"; volume: "6:16836. doi"; pages: " 
            10.1038/srep16836"
COMMENT     Sequence Label: R26TV LSL-TRL
FEATURES             Location/Qualifiers
     source          1..18186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..1086
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    join(1093..1158,1159..1196)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     enhancer        1210..1589
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1591..1864
                     /label=chicken beta-actin promoter
     intron          1865..2881
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2805..2827
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2889..2908
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2950..2983)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2990..3038
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3045..3848)
                     /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(3156..3175)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3766..3785
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     intron          4662..4727
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4857..4877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5302..5436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5339..5358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5393..5412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5456..5489)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             5526..6146
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(5599..5618)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             6147..6167
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6270..6464
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             6546..6602
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6603..7307
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     CDS             7323..7379
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7380..9032
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(7446..7464)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(9070..9086)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    9105..9329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    9417..13674
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     promoter        13723..14213
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     14146..14165
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             14240..14809
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphteria toxin Fragment A"
                     /label=DTA
                     /translation="DDVVILLIFCMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDD
                     DWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELG
                     LSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEIN
                     FETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          14886..14951
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    15025..15249
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(15341..15361)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(15341..15359)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(15380..15396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15380..15396)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15393..15415)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15404..15420
                     /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(15428..15434,15435..15452,15453..15458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15473..15494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15611..15628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15782..16370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15862..15881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16541..17332,17333..17401))
                     /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     17164..17183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17402..17506)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(17532..17987)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(17669..17690)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     17881..17900
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     18114..18136
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     18128..18145
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     18129..18145
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     18155..18174
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        18155..18173
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccccgcggca ggccctccga gcgtggtgga gccgttctgt gagacagccg ggtacgagtc
       61 gtgacgctgg aaggggcaag cgggtggtgg gcaggaatgc ggtccgccct gcagcaaccg
      121 gagggggagg gagaagggag cggaaaagtc tccaccggac gcggccatgg ctcggggggg
      181 ggggggcagc ggaggacgct tccggccgac gtctcgtcgc tgattggctt ttttcctccc
      241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
      301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
      361 ggtggggtga ggcgagctga cgtgcgggcg cggtcggcct ctggcggggc gggggagggg
      421 agggagggtc agcgaaagta gctcgcgcgc gagcggccgc ccaccctccc cttcctctgg
      481 gggagtcgtt ttacccgccg ccggccgggc ctcgtcgtct gattggctct cggggcccag
      541 aaaactggcc cttgccattg gctcgtgttc gtgcaagttg agtccatccg ccggccagcg
      601 ggggcggcga ggaggcgctc ccaggttccg gccctcccct cggccccgcg ccgcagagtc
      661 tggccgcgcg cccctgcgca acgtggcagg aagcgcgcgc tgggggcggg gacgggcagt
      721 agggctgagc ggctgcgggg cgggtgcaag cacgtttccg acttgagttg cctcaagagg
      781 ggcgtgctga gccagacctc catcgcgcac tccggggagt ggagggaagg agcgagggct
      841 cagttgggct gttttggagg caggaagcac ttgctctccc aaagtcgctc tgagttgtta
      901 tcagtaaggg agctgcagtg gagtaggcgg ggagaaggcc gcacccttct ccggaggggg
      961 gaggggagtg ttgcaatacc tttctgggag ttctctgctg cctcctggct tctgaggacc
     1021 gccctgggcc tgggagaatc ccttgccccc tcttcccctc gtgatctgca actccagtct
     1081 ttctagcatc tgtagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg
     1141 tccctttttt ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtggtt
     1201 aattaagtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     1261 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     1321 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     1381 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     1441 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     1501 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1561 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     1621 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     1681 gtgcagcgat gggggcgggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
     1741 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     1801 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     1861 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     1921 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     1981 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     2041 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
     2101 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
     2161 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     2221 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     2281 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     2341 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     2401 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     2461 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     2521 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     2581 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     2641 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     2701 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     2761 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     2821 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     2881 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga
     2941 gttaattaaa taacttcgta tagcatacat tatacgaagt tatggatccg aacaaacgac
     3001 ccaacacccg tgcgttttat tctgtctttt tattgccgat cccctcagaa gaactcgtca
     3061 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     3121 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     3181 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     3241 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     3301 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     3361 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     3421 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     3481 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     3541 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     3601 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     3661 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     3721 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     3781 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatggcc
     3841 gatcccatat tggctgcagg gtcgctcggt gttcgaggcc acacgcgtca ccttaatatg
     3901 cgaagtggac ctgggaccgc gccgccccga ctgcatctgc gtgttcgaat tcgccaatga
     3961 caagacgctg ggcggggttt gctcgacatt gggtggaaac attccaggcc tgggtggaga
     4021 ggctttttgc ttcctcttgc aaaaccacac tgctcgacat tgggtggaaa cattccaggc
     4081 ctgggtggag aggctttttg cttcctcttg caaaaccaca ctgctcgact agtgattaaa
     4141 gtcgactcgg ggacaccaaa tatggcgatc tcggcctttt cgtttcttgg agctgggaca
     4201 tgtttgccat cgatccatct accaccagaa cggccgttag atctgctgcc accgttgttt
     4261 ccaccgaaga aaccaccgtt gccgtaacca ccacgacggt tgttgctaaa gaagctgcca
     4321 ccgccacggc caccgttgta gccgccgttg ttgttattgt agttgctcat gttatttctg
     4381 gcacttcttg gttttcctct taagtgagga ggaacataac cattctcgtt gttgtcgttg
     4441 atgcttaaat tttgcacttg ttcgctcagt tcagccataa tatgaaatgc ttttcttgtt
     4501 gttcttacgg aataccactt gccacctatc accacaacta actttttccc gttcctccat
     4561 ctcttttata ttttttttct cgagggatct ttgtgaagga accttacttc tgtggtgtga
     4621 cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa
     4681 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg
     4741 gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag
     4801 aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa
     4861 aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga
     4921 gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa
     4981 aagctgcact gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc
     5041 ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg
     5101 ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta
     5161 ataaggaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt
     5221 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     5281 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5341 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5401 tccaaactca tcaatgtatc ttatcatgtc tggatctgac atggtaagta agcttataac
     5461 ttcgtatagc atacattata cgaagttatg ggctaactcg aacgctagct gtgcgatcgt
     5521 ttaccatggc tagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg
     5581 aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc
     5641 ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga
     5701 tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt
     5761 tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag
     5821 tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct
     5881 ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc
     5941 attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg
     6001 aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc
     6061 accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa
     6121 aacaacttaa atgtgaaagt gggtcgccaa aaaagaagag aaaggtcgac ggcggtggtg
     6181 ctttgtctcc tcagcactct gctgtcactc aaggaagtat catcaagaac aaggagggca
     6241 tggatgctaa gtcactaact gcctggtccc ggacactggt gaccttcaag gatgtatttg
     6301 tggacttcac cagggaggag tggaagctgc tggacactgc tcagcagatc gtgtacagaa
     6361 atgtgatgct ggagaactat aagaacctgg tttccttggg ttatcagctt actaagccag
     6421 atgtgatcct ccggttggag aagggagaag agccctggct ggtggagaga gaaattcacc
     6481 aagagaccca tcctgattca gagactgcat ttgaaatcaa atcatcagtt aaactgggca
     6541 gcggtgccac taacttcagc ttgttgaagc aggccggaga cgtcgaagag aacccgggtc
     6601 caatgtctag actggacaag agcaaagtca taaacggagc tctggaatta ctcaatggtg
     6661 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta
     6721 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc
     6781 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc
     6841 ggaacaacgc caagtcatac cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc
     6901 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc
     6961 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact
     7021 ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga
     7081 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc
     7141 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac
     7201 agctaaagtg cgaaagcggc gggccgaccg acgcccttga cgattttgac ttagacatgc
     7261 tcccagccga tgcccttgac gattttgacc ttgacatgct ccccgggaag ttaggatccg
     7321 gagccacgaa cttctctctg ttaaagcaag caggagacgt ggaagaaaac cccgggccca
     7381 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa
     7441 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
     7501 cttttacaga tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg
     7561 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
     7621 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
     7681 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt
     7741 cgcagcctac cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
     7801 aaaagctccc aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat
     7861 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
     7921 ttgtgccaga gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat
     7981 ctactggtct gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc
     8041 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
     8101 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
     8161 gagtcgtctt aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca
     8221 agattcaaag tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga
     8281 ttgacaaata cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta
     8341 aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg
     8401 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
     8461 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
     8521 cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt
     8581 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
     8641 gagacatagc ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc
     8701 tgattaagta caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac
     8761 accccaacat cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc
     8821 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
     8881 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
     8941 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
     9001 aggccaagaa gggcggaaag atcgccgtgt aaaaacgttg ttgtttgggg ttgaattact
     9061 ctcccggggg atccactagt tctagagcgg ccgatcagcc tcgactgtgc cttctagttg
     9121 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     9181 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     9241 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     9301 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     9361 gagatccact agttctagcc tcgaggctag agcggccgcc gcggatatcg aattcgctag
     9421 aagatgggcg ggagtcttct gggcaggctt aaaggctaac ctggtgtgtg ggcgttgtcc
     9481 tgcaggggaa ttgaacaggt gtaaaattgg agggacaaga cttcccacag attttcggtt
     9541 ttgtcgggaa gttttttaat aggggcaaat aggaaaatgg aggataggag tcatctgggg
     9601 tttatgcagc aaaactacag gtatattgct tgtatccgcc tcggagattt ccatgaggag
     9661 ataaagacat gtcacccgag tttatactct cctgcttaga tcctactaca gtatgaaata
     9721 cagtgtgcga ggtagactat gtaagcagat ttaatcattt taaagagccc agtacttcat
     9781 atccatttct cccgctcctt ctgcagcctt atcaaaaggt atttagaaca ctcattttag
     9841 ccccattttc atttattata ctggcttatc caacccctag acagagcatt ggcattttcc
     9901 ctttcctgat cttagaagtc tgatgactca tgaaaccaga cagattagtt acatacacca
     9961 caaatcgagg ctgtagctgg ggcctcaaca ctgcagttct tttataactc cttagtacac
    10021 tttttgttga tcctttgcct tgatccttaa ttttcagtgt ctatcacctc tcccgtcagg
    10081 tggtgttcca catttgggcc tattctcagt ccagggagtt ttacaacaat agatgtattg
    10141 agaatccaac ctaaagctta actttccact cccatgaatg cctctctcct ttttctccat
    10201 tataactgag ctataccatt aatggtttca ggtggatgtc tcctccccca atatacctga
    10261 tgtatctaca tattgccagg ctgatatttt aagacataaa ggtatatttc attattgagc
    10321 cacatggtat tgattactgc tactaaaatt ttgtcattgt acacatctgt aaaaggtggt
    10381 tccttttgga atgcaaagtt caggtgtttg ttgtctttcc tgacctaagg tcttgtgagc
    10441 ttgtattttt tctatttaag cagtgctttc tcttggactg gcttgactca tggcattcta
    10501 cacgttattg ctggtctaaa tgtgattttg ccaagcttct tcaggaccta taattttgct
    10561 tgacttgtag ccaaacacaa gtaaaatgat taagcaacaa atgtatttgt gaagcttggt
    10621 ttttaggttg ttgtgttgtg tgtgcttgtg ctctataata atactatcca ggggctggag
    10681 aggtggctcg gagttcaaga gcacagactg ctcttccaga agtcctgagt tcaattccca
    10741 gcaaccacat ggtggctcac aaccatctgt aatgggatct gatgccctct tctggtgtgt
    10801 ctgaagacca caagtgtatt cacattaaat aaataatcct ccttcttctt cttttttttt
    10861 ttttaaagag aatctgtctc cagtagaatt actgaagtaa tgaaatactt tgtgtttgtt
    10921 ccaatatgga gccaataatc aaatactctt agcactggaa atgtaccaag gaactatttt
    10981 atttaagtga ctgtggacag aggagccata actgcagact tgtgggatac agaagaccaa
    11041 tgcagactta atgtcttttc tcttacacta agcaataaag aaataaaaat tgaacttcta
    11101 gtatcctatt tgttaaactg ctagctttac taacttttgt gcttcatcta tacaaagctg
    11161 aaagctaagt ctgcagccat tactaaacat gaaagcaagt aatgataatt ttggatttca
    11221 aaaatgtagg gccagagttt agccagccag tggtggtgct tgcctttatg ccttaatccc
    11281 agcactctgg aggcagagac aggcagatct ctgagtttga gcccagcctg gtctacacat
    11341 caagttctat ctaggatagc caggaataca cacagaaacc ctgttgggga ggggggctct
    11401 gagatttcat aaaattataa ttgaagcatt ccctaatgag ccactatgga tgtggctaaa
    11461 tccgtctacc tttctgatga gatttgggta ttattttttc tgtctctgct gttggttggg
    11521 tcttttgaca ctgtgggctt tcttaaagcc tccttccctg ccatgtggtc tcttgtttgc
    11581 tactaacttc ccatggctta aatggcatgg ctttttgcct tctaagggca gctgctgagt
    11641 ttgcagcctg atttccaggg tggggttggg aaatctttca aacactaaaa ttgtccttta
    11701 attttttttt aaaaaatggg ttatataata aacctcataa aatagttatg aggagtgagg
    11761 tggactaata ttaatgagtc cctcccctat aaaagagcta ttaaggcttt ttgtcttata
    11821 ctaacttttt ttttaaatgt ggtatcttta gaaccaaggg tcttagagtt ttagtataca
    11881 gaaactgttg catcgcttaa tcagattttc tagtttcaaa tccagagaat ccaaattctt
    11941 cacagccaaa gtcaaattaa gaatttctga ctttaatgtt atttgctact gtgaatataa
    12001 aatgatagct tttcctgagg cagggtctca ctatgtatct ctgcctgatc tgcaacaaga
    12061 tatgtagact aaagttctgc ctgcttttgt ctcctgaata ctaaggttaa aatgtagtaa
    12121 tacttttgga acttgcaggt cagattcttt tataggggac acactaaggg agcttgggtg
    12181 atagttggta aatgtgttta agtgatgaaa acttgaatta ttatcaccgc aacctacttt
    12241 ttaaaaaaaa aagccaggcc tgttagagca tgctaaggga tccctaggac ttgctgagca
    12301 cacaagagta gtacttggca ggctcctggt gagagcatat ttcaaaaaac aaggcagaca
    12361 accaagaaac tacagtaagg ttacctgtct ttaaccatct gcatatacac agggatatta
    12421 aaatattcca aataatattt cattcaagtt ttcccccatc aaattgggac atggatttct
    12481 ccggtgaata ggcagagttg gaaactaaac aaatgttggt tttgtgattt gtgaaattgt
    12541 tttcaagtga tagttaaagc ccatgagata cagaacaaag ctgctatttc gaggtctctt
    12601 ggttatactc agaagcactt ctttgggttt ccctgcacta tcctgatcat gtgctaggcc
    12661 tccttaggct gattgttgtt caaataactt aagtttcctg tcaggtgatg tcatatgatt
    12721 tcatatatca aggcaaaaca tgttatatat gttaaacatt tgacttaatg tgaaagttag
    12781 gtctttgtgg gttttgattt taatttcaaa acctgagcta aataagtcat tttacatgtc
    12841 ttacatttgg tgaattgtat attgtggttt gcaggcaaga ctctctgacc tagtaaccct
    12901 cctatagagc actttgctgg gtcacaagtc taggagtcaa gcatttcacc ttgaagttga
    12961 gacgttttgt tagtgtatac tagttatatg ttggaggaca tgtttatcca gaagatattc
    13021 aggactattt ttgactgggc taaggaattg attctgatta gcactgttag tgagcattga
    13081 gtggccttta ggcttgaatt ggagtcactt gtatatctca aataatgctg gcctttttta
    13141 aaagcccttg ttctttatca ccctgttttc tacataattt ttgttcaaag aaatacttgt
    13201 ttggatctcc ttttgacaac aatagcatgt tttcaagcca tatttttttt cctttttttt
    13261 tttttttttg gtttttcgag acagggtttc tctgtatagc cctggctgtc ctggaactca
    13321 ctttgtagac caggctggcc tcgaactcag aaatccgcct gcctctgcct cctgagtgcc
    13381 gggattaaag gcgtgcacca ccacgcctgg ctaagttgga tattttgtat ataactataa
    13441 ccaatactaa ctccactggg tggattttta attcagtcag tagtcttaag tggtctttat
    13501 tggcccttat taaaatctac tgttcactct aacagaggct gttggactag tggactaagc
    13561 aacttcctac ggatatacta gcagataagg gtcagggata gaaactagtc tagcgttttg
    13621 tatacctacc agcttatact accttgttct gatagaaata tttaggacat ctagcttatc
    13681 gatccgtcga cggtatcgat aagcttgata tcgaattcta ccgggtaggg gaggcgcttt
    13741 tccaaggcag tctgagcatg cgcttagcag ccccgctggc acttggcgct acacaagtgg
    13801 ccttggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
    13861 ggccccttcg cgccaccttc tctcctcccc tagtcaggaa gttccccccc gccccgcagc
    13921 tcgcgtcgta ggacgtgaca aatggaagta gcacgtctca ctagtctcgt cagatggaca
    13981 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
    14041 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
    14101 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
    14161 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
    14221 ggtcctcgcc atggatcctg atgatgttgt tattcttcta atcttttgta tggaaaactt
    14281 ttcttcgtac cacgggacta aacctggtta tgtagattcc attcaaaaag gtatacaaaa
    14341 gccaaaatct ggtacacaag gaaattatga cgatgattgg aaagggtttt atagtaccga
    14401 caataaatac gacgctgcgg gatactctgt agataatgaa aacccgctct ctggaaaagc
    14461 tggaggcgtg gtcaaagtga cgtatccagg actgacgaag gttctcgcac taaaagtgga
    14521 taatgccgaa actattaaga aagagttagg tttaagtctc actgaaccgt tgatggagca
    14581 agtcggaacg gaagagttta tcaaaaggtt cggtgatggt gcttcgcgtg tagtgctcag
    14641 ccttcccttc gctgagggga gttctagcgt tgaatatatt aataactggg aacaggcgaa
    14701 agcgttaagc gtagaacttg agattaattt tgaaacccgt ggaaaacgtg gccaagatgc
    14761 gatgtatgag tatatggctc aagcctgtgc aggaaatcgt gtcaggcgat ctctttgtga
    14821 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
    14881 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
    14941 gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg aatgcagatc
    15001 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
    15061 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
    15121 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
    15181 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
    15241 gctctatggc ttctgaggcg gaaagaacca gctggggctc gacctcgagg gggggcccgc
    15301 ggccgccctg cagggcgatc gctacgtaca gcttttgttc cctttagtga gggttaattg
    15361 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    15421 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    15481 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    15541 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    15601 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    15661 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    15721 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    15781 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    15841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    15901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    15961 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    16021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    16081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    16141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    16201 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
    16261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    16321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    16381 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    16441 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    16501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    16561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    16621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    16681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    16741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    16801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    16861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    16921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    16981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    17041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    17101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    17161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    17221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    17281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    17341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    17401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    17461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    17521 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
    17581 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
    17641 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
    17701 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
    17761 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
    17821 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
    17881 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
    17941 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
    18001 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
    18061 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
    18121 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
    18181 ggagct
//
