LOCUS       sequence-481552-       13387 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Inducible YAP knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13387)
  AUTHORS   Er EE, Valiente M, Ganesh K, Zou Y, Agrawal S, Hu J, Griscom B, 
            Rosenblum M, Boire A, Brogi E, Giancotti FG, Schachner M, Malladi S,
            Massague J
  TITLE     Pericyte-like spreading by disseminated cancer cells activates YAP 
            and MRTF for metastatic colonization.
  JOURNAL   Nat Cell Biol. 2018 Aug;20(8):966-978. doi: 
            10.1038/s41556-018-0138-8. Epub 2018 Jul 23.
  PUBMED    30038252
REFERENCE   2  (bases 1 to 13387)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41556-018-0138-8"; journalName: "Nat Cell Biol"; date: 
            "2018-08"; volume: "20"; issue: "8"; pages: "966-978"
COMMENT     Sequence Label: TetON-shYAP
FEATURES             Location/Qualifiers
     source          1..13387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      134..143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(147..165)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(157..204)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    240..258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    275..293
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    311..329
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    347..365
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    382..400
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    418..436
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        448..486
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     483..507
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             523..1218
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     ncRNA           1231..1352
                     /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           1380..1394
                     /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"
     ncRNA           1422..1548
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        complement(1626..1644)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1655..2866
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2703..2722
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(2879..2912)
                     /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             2919..3626
                     /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(3627..3660)
                     /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    3683..4255
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3850..3867)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4077..4096
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4258..4860
                     /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     4757..4777
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4876..5464
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4929..4949)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5347..5358)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5671..5904
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6005..6022)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6011..6235
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6281..6709
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6368..6387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6578..6599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6718..6738)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6723..7052
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6903..7038
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6965..6984
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    8260..8381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8430..8446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8430..8446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8443..8465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8454..8470
                     /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(8478..8484,8485..8502,8503..8508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8523..8544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8661..8678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8832..9420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8912..8931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9591..10382,10383..10451))
                     /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     10214..10233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10452..10556)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10604..10738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             10906..11539
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11586..11711
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12208..12441
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12626..12670
                     /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    12968..13085
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(join(13150..13387,1..137))
                     /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"
ORIGIN
        1 acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga agtcccggga gaacccgagc
       61 cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg cggtgagcac cggaacggca
      121 ctggtcaact tggccatggt ggccctccta tagtgagtcg tattatacta tgccgatata
      181 ctatgccgat gattaattgt caacacgtgc tgcaggtccg aggttctaga cgagtttact
      241 ccctatcagt gatagagaac gatgtcgagt ttactcccta tcagtgatag agaacgtatg
      301 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      361 agagaacgta tgtcgagttt atccctatca gtgatagaga acgtatgtcg agtttactcc
      421 ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc
      481 agagctcgtt tagtgaaccg tcagatcgca ccggtcgcca ccatgagcga gctgatcaag
      541 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
      601 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgaagat caaggtggtc
      661 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc
      721 aaagccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccctgag
      781 ggcttcacat gggagagaat caccacatac gaagacgggg gcgtgctgac cgctacccag
      841 gacaccagct tccagaacgg ctgcatcatc tacaacgtca agatcaacgg ggtgaacttc
      901 ccatccaacg gccctgtgat gcagaagaaa acacgcggct gggaggccaa caccgagatg
      961 ctgtaccccg ctgacggcgg cctgagaggc cacagccaga tggccctgaa gctcgtgggc
     1021 gggggctacc tgcactgctc cttcaagacc acatacagat ccaagaaacc cgctaagaac
     1081 ctcaagatgc ccggcttcca cttcgtggac cacagactgg aaagaatcaa ggaggccgac
     1141 aaagagacct acgtcgagca gcacgagatg gctgtggcca agtactgcga cctccctagc
     1201 aaactggggc acagataatc gatagtttgt ttgaatgagg cttcagtact ttacagaatc
     1261 gttgcctgca catcttggaa acacttgctg ggattacttc ttcaggttaa cccaacagaa
     1321 ggctcgagaa ggtatattgc tgttgacagt gagcgcaggg ttcataacag gcataaatag
     1381 tgaagccaca gatgtattta tgcctgttat gaaccctttg cctactgcct cggaattcaa
     1441 ggggctactt taggagcaat tatcttgttt actaaaactg aataccttgc tatctctttg
     1501 atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt tcaattggaa
     1561 gacttccata ttgttgatgt ccacgcatgt ctaatgtgtg caagcggggc ggttagatgc
     1621 gctatcctat agtgagtcgt attaaattac gcgtggcctc cgcgccgggt tttggcgcct
     1681 cccgcgggcg cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag
     1741 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc
     1801 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca
     1861 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct
     1921 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg
     1981 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt
     2041 gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc
     2101 cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag
     2161 caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt
     2221 cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat
     2281 ggtgggcggc aagaacccaa ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg
     2341 ggtgagatgg gctggggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     2401 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     2461 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     2521 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     2581 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     2641 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     2701 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     2761 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagactag
     2821 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacgcta gcggatccat
     2881 aacttcgtat agtatacatt atacgaagtt atgccaccat gtctaggctg gacaagagca
     2941 aagtcataaa cggagctctg gaattactca atggtgtcgg tatcgaaggc ctgacgacaa
     3001 ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca
     3061 agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccacttct
     3121 gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag tcataccgct
     3181 gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga
     3241 aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg
     3301 agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg
     3361 aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc
     3421 cacttctgag acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt
     3481 tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc
     3541 cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgatt
     3601 ttgaccttga catgctcccc gggtaaataa cttcgtatag tatacattat acgaagttat
     3661 ggatccgcgg ccgcaaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     3721 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     3781 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     3841 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     3901 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     3961 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     4021 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     4081 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     4141 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     4201 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga taataccatg
     4261 gccaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     4321 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     4381 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4441 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4501 agcgtcgaag cgggggcggt gttcgccgag atcggctcgc gcatggccga gttgagcggt
     4561 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     4621 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4681 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ctggggtgcc cgccttcctg
     4741 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4801 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     4861 gttcgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4921 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4981 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5041 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5101 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5161 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5221 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     5281 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5341 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5401 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5461 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     5521 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5581 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5641 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
     5701 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5761 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5821 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5881 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     5941 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgttta aacccgctga
     6001 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6061 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6121 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6181 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     6241 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     6301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6421 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6481 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6541 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6601 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6661 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     6721 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6781 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6841 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6901 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6961 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     7021 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     7081 tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     7141 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     7201 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     7261 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     7321 ttccggaagt gcttgacatt ggggaattaa ttcagcgaga gcctgaccta ttgcatctcc
     7381 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     7441 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     7501 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     7561 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     7621 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     7681 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     7741 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     7801 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     7861 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     7921 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     7981 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     8041 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     8101 cccagcactc gtccgagggc aaaggaatag cacgtgctac gagatttcga ttccaccgcc
     8161 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     8221 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     8281 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8341 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     8401 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     8461 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     8521 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     8581 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     8641 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     8701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     9001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     9061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     9121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     9181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     9241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     9301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     9361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     9421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     9481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    10021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    10081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    10141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    10201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    10261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    10321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    10381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    10441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10561 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc
    10621 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    10681 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
    10741 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
    10801 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
    10861 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttggaa gggctaattc
    10921 actcccaaag aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
    10981 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
    11041 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
    11101 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
    11161 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
    11221 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
    11281 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
    11341 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    11401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    11461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    11521 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
    11581 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
    11641 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
    11701 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
    11761 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
    11821 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
    11881 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
    11941 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
    12001 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccggccgct
    12061 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
    12121 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
    12181 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
    12241 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
    12301 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
    12361 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
    12421 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
    12481 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
    12541 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
    12601 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
    12661 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
    12721 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
    12781 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
    12841 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
    12901 tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca
    12961 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
    13021 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
    13081 attttcgggt ttattacagg gacagcagag atccagtttg gttagtaccg ggcccgctct
    13141 agtccggaat cagtcctgct cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg
    13201 cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg gcccggaggc
    13261 gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt ccaggccgcg
    13321 cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg cgctgatgaa
    13381 cagggtc
//
