LOCUS       sequence-460030-       14924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression, inducible PBRM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14924)
  AUTHORS   Gao W, Li W, Xiao T, Liu XS, Kaelin WG Jr
  TITLE     Inactivation of the PBRM1 tumor suppressor gene amplifies the 
            HIF-response in VHL-/- clear cell renal carcinoma.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):1027-1032. doi: 
            10.1073/pnas.1619726114. Epub 2017 Jan 12.
  PUBMED    28082722
REFERENCE   2  (bases 1 to 14924)
  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"; doi: 
            "10.1073/pnas.1619726114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-01-31- 31"; volume: "114"; issue: "5"; pages: 
            "1027-1032"
COMMENT     Sequence Label: WT PBRM1- TRIPZ-neo
FEATURES             Location/Qualifiers
     source          1..14924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     814..833
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(990..1023)
                     /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             1030..1737
                     /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(1738..1771)
                     /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    1791..2364
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1958..1975)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2185..2204
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2365..3165
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GSMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     LTR             3237..3470
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3542..3663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3579..3598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3633..3652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3703..3838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3858..3877)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3859..3877)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(3900..4258)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3940..3961)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4152..4171
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4285..4389
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4391..4748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4391..4582
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4414..4434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4599..4734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4783..5577
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4783..5577
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5765..5784)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5809..5856
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6959..7262
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7264..7462
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             7480..7660
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7707..7832
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8329..8562
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8747..8791
                     /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    9089..9206
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        9280..9594
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    9288..9306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9324..9342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9359..9377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9395..9413
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9431..9449
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9466..9484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9502..9520
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     9567..9591
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             9638..9661
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9662..9688
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             14615..14680
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        join(14690..14924,1..977)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
       61 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
      121 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
      181 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
      241 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
      301 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
      361 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
      421 gccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctggggca ccatctgggg
      481 accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg
      541 gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc
      601 gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag
      661 gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc
      721 tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt
      781 ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct
      841 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg
      901 ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc tggccgtttt
      961 tggctttttt gttagacgct agcggatcca taacttcgta tagtatacat tatacgaagt
     1021 tatgccacca tgtctaggct ggacaagagc aaagtcataa acggagctct ggaattactc
     1081 aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     1141 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     1201 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     1261 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct
     1321 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     1381 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     1441 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     1501 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     1561 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     1621 gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta
     1681 gacatgctcc cagccgatgc ccttgacgat tttgaccttg acatgctccc cgggtaaata
     1741 acttcgtata gtatacatta tacgaagtta tggatccgcg gccgcatccg cccctctccc
     1801 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1861 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1921 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     1981 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     2041 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     2101 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     2161 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     2221 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2281 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2341 ttcctttgaa aaacacgatg ataaggatcc atgattgaac aagatggatt gcacgcaggt
     2401 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2461 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2521 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2581 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2641 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     2701 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     2761 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     2821 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     2881 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     2941 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3001 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3061 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3121 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaggtac ctttaagacc
     3181 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     3241 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     3301 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     3361 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     3421 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     3481 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     3541 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3661 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     3721 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3781 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3841 gggacgtacc caattcgccc tatagtgagt cgtattagca tgtgagcaaa aggccagcat
     3901 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3961 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4021 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4081 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4141 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4201 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4381 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4681 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4741 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5761 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5821 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5881 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5941 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6001 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctggc cttttgctca
     6841 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     6901 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     6961 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7021 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7081 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7141 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7201 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7261 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     7321 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     7381 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     7441 ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag
     7501 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     7561 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7621 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     7681 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     7741 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     7801 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     7861 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     7921 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     7981 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     8041 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     8101 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     8161 cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     8221 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     8281 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     8341 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     8401 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     8461 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     8521 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     8581 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     8641 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     8701 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     8761 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     8821 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     8881 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     8941 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     9001 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
     9061 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     9121 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     9181 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     9241 ggttagtacc gggcccctga ggccctttcg tcttcattcg agtttactcc ctatcagtga
     9301 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc
     9361 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
     9421 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata
     9481 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt
     9541 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     9601 atctagactg ccggatcgaa ttgtcgacga attcatggac tacaaagacg atgacgacaa
     9661 gtacccatac gatgttccag attacgctgg ttccaagaga agaagagcta cctccccttc
     9721 cagcagtgtc agcggggact ttgatgatgg gcaccattct gtgtcaacac caggcccaag
     9781 caggaaaagg aggagacttt ccaatcttcc aactgtagat cctattgccg tgtgccatga
     9841 actctataat accatccgag actataagga tgaacagggc agacttctct gtgagctctt
     9901 cattagggca ccaaagcgaa gaaatcaacc agactattat gaagtggttt ctcagcccat
     9961 tgacttgatg aaaatccaac agaaactaaa aatggaagag tatgatgatg ttaatttgct
    10021 gactgctgac ttccagcttc tttttaacaa tgcaaagtcc tattataagc cagattctcc
    10081 tgaatataaa gccgcttgca aactctggga tttgtacctt cgaacaagaa atgagtttgt
    10141 tcagaaagga gaagcagatg acgaagatga tgatgaagat gggcaagaca atcagggcac
    10201 agtgactgaa ggatcttctc cagcttactt gaaggagatc ctggagcagc ttcttgaagc
    10261 catagttgta gctacaaatc catcaggacg tctcattagc gaactttttc agaaactgcc
    10321 ttctaaagtg caatatccag attattatgc aataattaag gagcctatag atctcaagac
    10381 cattgcccag aggatacaga atggaagcta caaaagtatt catgcaatgg ccaaagatat
    10441 agatctcctc gcaaaaaatg ccaaaactta taatgagcct ggctctcaag tattcaagga
    10501 tgcaaattca attaaaaaaa tattttatat gaaaaaggct gaaattgaac atcatgaaat
    10561 ggctaagtca agtcttcgaa tgaggactcc atccaacttg gctgcagcca gactgacagg
    10621 tccttcacac agtaaaggca gccttggtga agagagaaat cccactagca agtattaccg
    10681 taataaaaga gcagtacaag gaggtcgttt atcagcaatt acaatggcac ttcaatatgg
    10741 ctcagaaagt gaagaagatg ctgctttagc tgctgcacgc tatgaagagg gagagtcaga
    10801 agcagaaagc atcacttcct ttatggatgt ttcaaatcct ttttatcagc tttatgacac
    10861 agttaggagt tgtcggaata accaagggca gctaatagct gaaccttttt accatttgcc
    10921 ttcaaagaaa aaataccctg attattacca gcaaattaaa atgcccatat cactacaaca
    10981 gatccgaaca aaactgaaga atcaagaata tgaaacttta gatcatttgg agtgtgatct
    11041 gaatttaatg tttgaaaatg ccaaacgcta taatgtgccc aattcagcca tctacaagcg
    11101 agttctaaaa ttgcagcaag ttatgcaggc aaagaagaaa gagcttgcca ggagagacga
    11161 tatcgaggac ggagacagca tgatctcttc agccacctct gatactggta gtgccaaaag
    11221 aaaaagtaaa aagaacataa gaaagcagcg aatgaaaatc ttattcaatg ttgttcttga
    11281 agctcgagag ccaggttcag gcagaagact ttgtgaccta tttatggtta aaccatccaa
    11341 aaaggactat cctgattatt ataaaatcat cttggagcca atggacttga aaataattga
    11401 gcataacatc cgcaatgaca aatatgctgg tgaagaggga atgatagaag acatgaagct
    11461 gatgttccgg aatgccaggc actataatga ggagggctcc caggtttata atgatgcaca
    11521 tatcctggag aagttactca aggagaaaag gaaagagctg ggcccactgc ctgatgatga
    11581 tgacatggct tctcccaaac tcaagctgag taggaagagt ggcatttctc ctaaaaaatc
    11641 aaaatacatg actccaatgc agcagaaact aaatgaggtc tatgaagctg taaagaacta
    11701 tactgataag aggggtcgcc gcctcagtgc catatttctg aggcttccct ctagatctga
    11761 gttgcctgac tactatctga ctattaaaaa gcccatggac atggaaaaaa ttcgaagtca
    11821 catgatggcc aacaagtacc aagatattga ctctatggtt gaggactttg tcatgatgtt
    11881 taataatgcc tgtacgtaca atgagccgga gtctttgatc tacaaagatg ctcttgttct
    11941 acacaaagtc ctgcttgaaa cacgcagaga cctggaggga gatgaggact ctcatgtccc
    12001 aaatgtgact ttgctgattc aagagcttat ccacaatctt tttgtgtcag tcatgagtca
    12061 tcaggatgat gagggaagat gctacagcga ttctttagca gaaattcctg ctgtggatcc
    12121 caactttcct aacaaaccac cccttacatt tgacataatt aggaagaatg ttgaaaataa
    12181 tcgctaccgt cggcttgatt tatttcaaga gcatatgttt gaagtattgg aacgagcaag
    12241 aaggatgaat cggacagatt cagaaatata tgaagatgca gtagaacttc agcagttttt
    12301 tattaaaatt cgtgatgaac tctgcaaaaa tggagagatt cttctttcac cggcactcag
    12361 ctataccaca aaacatttgc ataatgatgt ggagaaagag agaaaggaaa aattgccaaa
    12421 agaaatagag gaagataaac taaaacgaga agaagaaaaa agagaagctg aaaagagtga
    12481 agattcctct ggtgctgcag gcctctcagg cttacatcgc acatacagcc aggactgtag
    12541 ctttaaaaac agcatgtacc atgttggaga ttacgtctat gtggaacctg cagaggccaa
    12601 cctacaacca catatcgtct gtattgaaag actgtgggag gattcagctg gtgaaaaatg
    12661 gttgtatggc tgttggtttt accgaccaaa tgaaacattc cacctggcta cacgaaaatt
    12721 tctagaaaaa gaagttttta agagtgacta ttacaacaaa gttccagtta gtaaaattct
    12781 aggcaagtgt gtggtcatgt ttgtcaagga atactttaag ttatgcccag aaaacttccg
    12841 agatgaggat gtttttgtct gtgaatcacg gtattctgcc aaaaccaaat cttttaagaa
    12901 aattaaactg tggaccatgc ccatcagctc agtcaggttt gtccctcggg atgtgcctct
    12961 gcctgtggtt cgcgtggcct ctgtatttgc aaatgcagat aaaggtgatg atgagaagaa
    13021 tacagacaac tcagaggaca gtcgagctga agacaatttt aacttggaaa aggaaaaaga
    13081 agatgtccct gtggaaatgt ccaatggtga accaggttgc cactactttg agcagctcca
    13141 ttacaatgac atgtggctga aggttggcga ctgtgtcttc atcaagtccc atggcctggt
    13201 gcgtcctcgt gtgggcagaa ttgaaaaagt atgggttcga gatggagctg catattttta
    13261 tggccccatc ttcattcacc ctgaagaaac agagcatgag cccacaaaaa tgttctacaa
    13321 aaaagaagta tttctgagta atctggaaga aacctgcccc atgacatgta ttctcggaaa
    13381 gtgtgctgtg ttgtcattca aggacttcct ctcctgcagg ccaactgaaa taccagaaaa
    13441 tgacattctg ctttgtgaga gccgctacaa tgagagcgac aagcagatga agaaattcaa
    13501 aggattgaag aggttttcac tctctgctaa agtggtagat gatgaaattt actacttcag
    13561 aaaaccaatt gttcctcaga aggagccatc acctttgctg gaaaagaaga tccagttgct
    13621 agaagctaaa tttgccgagt tagaaggtgg agatgatgat attgaagaga tgggagaaga
    13681 agatagtgag gtcattgaac ctccttctct acctcagctt cagacccccc tggccagtga
    13741 gctggacctc atgccctaca cacccccaca gtctacccca aagtctgcca aaggcagtgc
    13801 aaagaaggaa ggctccaaac ggaaaatcaa catgagtggc tacatcctgt tcagcagtga
    13861 gatgagggct gtgattaagg cccaacaccc agactactct ttcggggagc tcagccgcct
    13921 ggtggggaca gaatggagaa atcttgagac agccaagaaa gcagaatatg aaggcatgat
    13981 gggtggctat ccgccaggcc ttccaccttt gcagggccca gttgatggcc ttgttagcat
    14041 gggcagcatg cagccacttc accctggggg gcctccaccc caccatcttc cgccaggtgt
    14101 gcctggcctc ccgggcatcc caccaccggg tgtgatgaac caaggagtgg cccctatggt
    14161 agggactcca gcaccgggtg gaagtccata tggacaacag gtgggagttt tggggcctcc
    14221 agggcagcag gcaccacctc catatcccgg cccacatcca gctggacccc ctgtcataca
    14281 gcagccaaca acacccatgt ttgtagctcc cccaccaaag acccagcggc ttcttcactc
    14341 agaggcctac ctgaaataca ttgaaggact cagtgcggag tccaacagca ttagcaagtg
    14401 ggatcagaca ctggcagctc gaagacgcga cgtccatttg tcgaaagaac aggagagccg
    14461 cctaccctct cactggctga aaagcaaagg ggcccacacc accatggcag atgccctctg
    14521 gcgccttcga gatttgatgc tccgggacac cctcaacatt cgccaagcat acaacctaga
    14581 aaatgtttaa ttaaggccgg ccctcgaggg aggtgactac aaagaccatg acggtgatta
    14641 taaagatcat gacatcgatt acaaggatga cgatgacaag tagacgcgtg gcctccgcgc
    14701 cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac
    14761 gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc
    14821 tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg
    14881 ggtgactcta gggcactggt tttctttcca gagagcggaa cagg
//
