LOCUS       sequence-433431-       13171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox inducible shRNA targeting WT-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13171)
  AUTHORS   Potluri S, Assi SA, Chin PS, Coleman DJL, Pickin A, Moriya S, Seki 
            N, Heidenreich O, Cockerill PN, Bonifer C
  TITLE     Isoform-specific and signaling-dependent propagation of acute 
            myeloid leukemia by Wilms tumor 1.
  JOURNAL   Cell Rep. 2021 Apr 20;35(3):109010. doi: 
            10.1016/j.celrep.2021.109010.
  PUBMED    33882316
REFERENCE   2  (bases 1 to 13171)
  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: "Cell Rep.";
            date: "2021-04-20"; volume: "35(3):109010. doi"; pages: " 
            10.1016/j.celrep.2021.109010"
COMMENT     Sequence Label: pTRIPZ-shWT1_B
FEATURES             Location/Qualifiers
     source          1..13171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(392..403)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             716..949
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1050..1067)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1056..1280
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1326..1754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1413..1432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1623..1644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1763..1783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1768..2097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1948..2083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2010..2029
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3305..3426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3475..3491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3475..3491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3488..3510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3499..3515
                     /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(3523..3529,3530..3547,3548..3553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3568..3589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3877..4465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3957..3976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4636..5427,5428..5496))
                     /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     5259..5278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5497..5601)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5649..5783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             5951..6584
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6631..6756
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7253..7486
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7671..7715
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8013..8130
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(8195..8569)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      8566..8575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8579..8597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8589..8636)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    8672..8690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8707..8725
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8743..8761
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8779..8797
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8814..8832
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8850..8868
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8880..8918
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8915..8939
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             8955..9650
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     ncRNA           9663..9784
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           9812..9826
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        9871..11082
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     10919..10938
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(11095..11128)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             11135..11842
                     /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"
     protein_bind    complement(11843..11876)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    11899..12471
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(12066..12083)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     12293..12312
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             12474..13076
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     12973..12993
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    join(13092..13171,1..509)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13145..13165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
       61 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
      121 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
      181 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
      241 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
      301 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
      361 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
      421 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
      481 tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa
      541 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
      601 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
      661 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa
      721 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
      781 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      841 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      901 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
      961 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     1021 ccttgacatt gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1081 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1141 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1201 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1261 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     1321 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1381 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1441 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1501 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1561 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1621 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1681 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1741 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1801 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     1861 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1921 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1981 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2041 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2101 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgatgaa aaagcctgaa
     2161 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     2221 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     2281 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     2341 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attaattcag
     2401 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     2461 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     2521 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     2581 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     2641 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     2701 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     2761 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     2821 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     2881 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     2941 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     3001 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     3061 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     3121 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     3181 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3241 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3301 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3361 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3421 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     5641 gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5761 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
     5821 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
     5881 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
     5941 acttagtagt tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg
     6001 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc
     6061 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta
     6121 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg
     6181 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac
     6241 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag
     6301 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga
     6361 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     6421 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6481 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6541 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     6601 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     6661 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     6721 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     6781 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     6841 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     6901 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     6961 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     7021 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     7081 accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     7141 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     7201 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     7261 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     7321 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     7381 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     7441 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     7501 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     7561 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     7621 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     7681 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     7741 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     7801 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     7861 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     7921 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     7981 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     8041 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     8101 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     8161 gtttggttag taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt
     8221 gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga
     8281 tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact
     8341 cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca
     8401 cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt
     8461 cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg
     8521 cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc
     8581 tatagtgagt cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt
     8641 gctgcaggtc cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga
     8701 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
     8761 aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat
     8821 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag
     8881 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
     8941 caccggtcgc caccatgagc gagctgatca aggagaacat gcacatgaag ctgtacatgg
     9001 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
     9061 agggcaccca gaccatgaag atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     9121 acatcctggc taccagcttc atgtacggca gcaaagcctt catcaaccac acccagggca
     9181 tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga atcaccacat
     9241 acgaagacgg gggcgtgctg accgctaccc aggacaccag cttccagaac ggctgcatca
     9301 tctacaacgt caagatcaac ggggtgaact tcccatccaa cggccctgtg atgcagaaga
     9361 aaacacgcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctgagag
     9421 gccacagcca gatggccctg aagctcgtgg gcgggggcta cctgcactgc tccttcaaga
     9481 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcttc cacttcgtgg
     9541 accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgaga
     9601 tggctgtggc caagtactgc gacctcccta gcaaactggg gcacagataa tcgatagttt
     9661 gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc
     9721 tgggattact tcttcaggtt aacccaacag aaggctcgag aaggtatatt gctgttgaca
     9781 gtgagcgagc atctgagacc agtgagaaat agtgaagcca cagatgtatt tctcactggt
     9841 ctcagatgcc tgcctactgc ctcgacgcgt ggcctccgcg ccgggttttg gcgcctcccg
     9901 cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc
     9961 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
    10021 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
    10081 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
    10141 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
    10201 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
    10261 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
    10321 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
    10381 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
    10441 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
    10501 ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg
    10561 agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
    10621 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
    10681 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
    10741 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
    10801 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
    10861 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
    10921 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
    10981 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa
    11041 ttgtccgcta aattctggcc gtttttggct tttttgttag acgctagcgg atccataact
    11101 tcgtatagta tacattatac gaagttatgc caccatgtct aggctggaca agagcaaagt
    11161 cataaacgga gctctggaat tactcaatgg tgtcggtatc gaaggcctga cgacaaggaa
    11221 actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga agaacaagcg
    11281 ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc acttctgccc
    11341 cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat accgctgtgc
    11401 tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa cagagaaaca
    11461 gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct ccctggagaa
    11521 cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat tggaggaaca
    11581 ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta tgcccccact
    11641 tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct tccttttcgg
    11701 cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg gcgggccgac
    11761 cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg acgattttga
    11821 ccttgacatg ctccccgggt aaataacttc gtatagtata cattatacga agttatggat
    11881 ccgcggccgc aaattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
    11941 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
    12001 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
    12061 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
    12121 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
    12181 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
    12241 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
    12301 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
    12361 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
    12421 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggcca
    12481 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
    12541 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
    12601 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
    12661 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
    12721 tcgaagcggg ggcggtgttc gccgagatcg gctcgcgcat ggccgagttg agcggttccc
    12781 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
    12841 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
    12901 ccgtcgtgct ccccggagtg gaggcggccg agcgcgctgg ggtgcccgcc ttcctggaga
    12961 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
    13021 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagttc
    13081 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
    13141 actatgttgc tccttttacg ctatgtggat a
//
