LOCUS       sequence-180587-       13405 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector expressing a doxycycline-inducible shRNA against 
            human CUX1 sequence starting at 5150 of M74099.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13405)
  AUTHORS   Ramdzan ZM, Vadnais C, Pal R, Vandal G, Cadieux C, Leduy L, Davoudi 
            S, Hulea L, Yao L, Karnezis AN, Paquet M, Dankort D, Nepveu A
  TITLE     RAS transformation requires CUX1-dependent repair of oxidative DNA 
            damage.
  JOURNAL   PLoS Biol. 2014 Mar 11;12(3):e1001807. doi: 
            10.1371/journal.pbio.1001807. eCollection 2014 Mar.
  PUBMED    24618719
REFERENCE   2  (bases 1 to 13405)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Biol."; date: "2014-03-11"; volume: "12(3):e1001807. doi"; pages: " 
            10.1371/journal.pbio.1001807. eCollection 2014 Mar"
COMMENT     Sequence Label: pTRIPZ-DoxOn-shCUX1-5150
FEATURES             Location/Qualifiers
     source          1..13405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2965..2989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3005..3700
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     ncRNA           3713..3833
                     /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           3861..3875
                     /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           3903..4029
                     /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(4124..4142)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4155..5366
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5203..5222
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(5379..5412)
                     /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             5419..6126
                     /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(6127..6160)
                     /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    6183..6755
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6350..6367)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6577..6596
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6758..7360
                     /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     7257..7277
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7376..7964
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7429..7449)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7847..7858)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8171..8404
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8505..8522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8511..8735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8781..9209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8868..8887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9078..9099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9218..9238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9223..9552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9403..9538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9465..9484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10760..10881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10930..10946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10930..10946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10943..10965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10954..10970
                     /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(10978..10984,10985..11002,11003..11008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11023..11044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11161..11178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11332..11920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11412..11431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12091..12882,12883..12951))
                     /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     12714..12733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12952..13056)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13104..13238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggtcgc
     3001 caccatgagc gagctgatca aggagaacat gcacatgaag ctgtacatgg agggcaccgt
     3061 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
     3121 gaccatgaag atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     3181 taccagcttc atgtacggca gcaaagcctt catcaaccac acccagggca tccccgactt
     3241 ctttaagcag tccttccctg agggcttcac atgggagaga atcaccacat acgaagacgg
     3301 gggcgtgctg accgctaccc aggacaccag cttccagaac ggctgcatca tctacaacgt
     3361 caagatcaac ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacacgcgg
     3421 ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctgagag gccacagcca
     3481 gatggccctg aagctcgtgg gcgggggcta cctgcactgc tccttcaaga ccacatacag
     3541 atccaagaaa cccgctaaga acctcaagat gcccggcttc cacttcgtgg accacagact
     3601 ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgaga tggctgtggc
     3661 caagtactgc gacctcccta gcaaactggg gcacagataa tcgatagttt gtttgaatga
     3721 ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc tgggattact
     3781 tcttcaggtt aacccaacag aaggctcgag aaggtatatg ctgttgacag tgagcgtcag
     3841 agcgataata cactattata gtgaagccac agatgtataa tagtgtatta tcgctctgat
     3901 gcctactgcc tcggaattca aggggctact ttaggagcaa ttatcttgtt tactaaaact
     3961 gaataccttg ctatctcttt gatacatttt tacaaagctg aattaaaatg gtataaatta
     4021 aatcactttt ttcaattgga agactaatgc ggccggccat tactccgtct cgtgtcttgt
     4081 tgcatatgtc tgctggtttg tttgatgttg tttgcgggcg ggccctatag tgagtcgtat
     4141 tacctaggac gcgtggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct
     4201 cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg
     4261 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     4321 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     4381 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     4441 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     4501 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     4561 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     4621 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     4681 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     4741 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     4801 ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac
     4861 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     4921 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     4981 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     5041 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     5101 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     5161 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     5221 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     5281 aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct
     5341 ggccgttttt ggcttttttg ttagacgcta gcggatccat aacttcgtat agtatacatt
     5401 atacgaagtt atgccaccat gtctaggctg gacaagagca aagtcataaa cggagctctg
     5461 gaattactca atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     5521 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     5581 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     5641 tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc
     5701 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     5761 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     5821 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     5881 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     5941 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     6001 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat
     6061 tttgacttag acatgctccc agccgatgcc cttgacgatt ttgaccttga catgctcccc
     6121 gggtaaataa cttcgtatag tatacattat acgaagttat ggatccgcgg ccgcaaattc
     6181 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     6241 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     6301 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     6361 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     6421 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     6481 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     6541 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     6601 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     6661 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     6721 ggggacgtgg ttttcctttg aaaaacacga taataccatg gccaccgagt acaagcccac
     6781 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     6841 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     6901 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     6961 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7021 gttcgccgag atcggctcgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7081 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7141 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7201 agtggaggcg gccgagcgcg ctggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7261 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7321 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gttcgcgtct ggaacaatca
     7381 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     7441 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     7501 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     7561 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     7621 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     7681 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     7741 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     7801 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     7861 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     7921 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc
     7981 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     8041 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     8101 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     8161 aagaggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     8221 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     8281 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     8341 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     8401 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     8461 tcagagagtg agaggccttg acattgttta aacccgctga tcagcctcga ctgtgccttc
     8521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     8581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     8641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     8701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     8761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     8821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     8881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     8941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     9001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     9061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     9121 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     9181 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     9241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     9301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     9361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     9421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     9481 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     9541 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     9601 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     9661 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     9721 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     9781 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     9841 ggggaattaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     9901 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     9961 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
    10021 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
    10081 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
    10141 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
    10201 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
    10261 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
    10321 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
    10381 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
    10441 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
    10501 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
    10561 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
    10621 aaaggaatag cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
    10681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
    10741 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
    10801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    10861 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    10921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    10981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    11041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    11101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
    11161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    11221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    11281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    11341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    11401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    11461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    11521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    11581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    11641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    11701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    11761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    11821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    11881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    11941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    12001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    12061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    12121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    12181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    12241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    12301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    12361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    12421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    12481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    12541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    12601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    12661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    12721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    12781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    12841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    12901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    12961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    13021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    13081 tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa
    13141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
    13201 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc
    13261 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc
    13321 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt
    13381 aaatatgtac tacaaactta gtagt
//
