LOCUS       sequence-210768-       13656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of gRNA resistant PBRM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13656)
  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 13656)
  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: pLenti-PGK-Hygro-amp-sgRNA-resistant PBRM1
FEATURES             Location/Qualifiers
     source          1..13656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    264..381
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        427..937
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     856..874
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    992..1016
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1049..1075
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(6022..6046)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6077..6665
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6130..6150)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6548..6559)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6721..7220
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6756..6779)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7153..7172
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7238..8263
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7238..8263
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             8637..8870
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8942..9063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8979..8998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9033..9052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9103..9238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9258..9277)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9259..9277)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9287..9304)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9287..9303)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9296..9318)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9445..9900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9532..9551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9742..9763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9926..10030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10031..10099,10100..10891)
                     /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     complement(10249..10268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11062..11650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11551..11570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11804..11821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11938..11959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11974..11979,11980..11997,11998..12004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12012..12028
                     /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)."
     primer_bind     12017..12039
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12036..12052
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12036..12052
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12071..12091
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12073..12091
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        12119..12345
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             12346..12526
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12573..12698
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    13191..13424
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13609..13653
                     /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"
ORIGIN
        1 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
       61 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
      121 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
      181 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
      241 ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca
      301 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
      361 attacaaaaa ttcaaaattt tatcgatcac gagactagcc tcgagaagct tgatatcgaa
      421 ttccacgggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
      481 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
      541 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
      601 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
      661 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
      721 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
      781 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
      841 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
      901 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg ttcgtcgact
      961 agtccagtgt ggtggaattc tgcagatatc aacaagtttg tacaaaaaag caggctttaa
     1021 aggaaccaat tcagtcgaca agcttatgta cccatacgat gttccagatt acgctggtac
     1081 cggttccaag agaagaagag ctacctcccc ttccagcagt gtcagcgggg actttgatga
     1141 tgggcaccat tctgtgtcaa caccaggccc aagcaggaaa aggaggagac tttccaatct
     1201 tccaactgta gatcctattg ccgtgtgcca tgaactctat aataccatcc gagactataa
     1261 ggatgaacag ggcagacttc tctgtgagct cttcattagg gcaccaaagc gaagaaatca
     1321 accagactat tatgaagtgg tttctcagcc cattgacttg atgaaaatcc aacagaaact
     1381 aaaaatggaa gagtatgatg atgttaattt gctgactgct gacttccagc ttctttttaa
     1441 caatgcaaag tcctattata agccagattc tcctgaatat aaagccgctt gcaaactctg
     1501 ggatttgtac cttcgaacaa gaaatgagtt tgttcagaaa ggagaagcag atgacgaaga
     1561 tgatgatgaa gatgggcaag acaatcaggg cacagtgact gaaggatctt ctccagctta
     1621 cttgaaggag atcctggagc agcttcttga agccatagtt gtagctacaa atccatcagg
     1681 acgtctcatt agcgaacttt ttcagaaact gccttctaaa gtgcaatatc cagattatta
     1741 tgcaataatt aaggagccta tagatctcaa gaccattgcc cagaggatac agaatggaag
     1801 ctacaaaagt attcatgcaa tggcgaaaga catcgacctg ctcgcaaaaa atgccaaaac
     1861 ttataatgag cctggctctc aagtattcaa ggatgcaaat tcaattaaaa aaatatttta
     1921 tatgaaaaag gctgaaattg aacatcatga aatggctaag tcaagtcttc gaatgaggac
     1981 tccatccaac ttggctgcag ccagactgac aggtccttca cacagtaaag gcagccttgg
     2041 tgaagagaga aatcccacta gcaagtatta ccgtaataaa agagcagtac aaggaggtcg
     2101 tttatcagca attacaatgg cacttcaata tggctcagaa agtgaagaag atgctgcttt
     2161 agctgctgca cgctatgaag agggagagtc agaagcagaa agcatcactt cctttatgga
     2221 tgtttcaaat cctttttatc agctttatga cacagttagg agttgtcgga ataaccaagg
     2281 gcagctaata gctgaacctt tttaccattt gccttcaaag aaaaaatacc ctgattatta
     2341 ccagcaaatt aaaatgccca tatcactaca acagatccga acaaaactga agaatcaaga
     2401 atatgaaact ttagatcatt tggagtgtga tctgaattta atgtttgaaa atgccaaacg
     2461 ctataatgtg cccaattcag ccatctacaa gcgagttcta aaattgcagc aagttatgca
     2521 ggcaaagaag aaagagcttg ccaggagaga cgatatcgag gacggagaca gcatgatctc
     2581 ttcagccacc tctgatactg gtagtgccaa aagaaaaagt aaaaagaaca taagaaagca
     2641 gcgaatgaaa atcttattca atgttgttct tgaagctcga gagccaggtt caggcagaag
     2701 actttgtgac ctatttatgg ttaaaccatc caaaaaggac tatcctgatt attataaaat
     2761 catcttggag ccaatggact tgaaaataat tgagcataac atccgcaatg acaaatatgc
     2821 tggtgaagag ggaatgatag aagacatgaa gctgatgttc cggaatgcca ggcactataa
     2881 tgaggagggc tcccaggttt ataatgatgc acatatcctg gagaagttac tcaaggagaa
     2941 aaggaaagag ctgggcccac tgcctgatga tgatgacatg gcttctccca aactcaagct
     3001 gagtaggaag agtggcattt ctcctaaaaa atcaaaatac atgactccaa tgcagcagaa
     3061 actaaatgag gtctatgaag ctgtaaagaa ctatactgat aagaggggtc gccgcctcag
     3121 tgccatattt ctgaggcttc cctctagatc tgagttgcct gactactatc tgactattaa
     3181 aaagcccatg gacatggaaa aaattcgaag tcacatgatg gccaacaagt accaagatat
     3241 tgactctatg gttgaggact ttgtcatgat gtttaataat gcctgtacgt acaatgagcc
     3301 ggagtctttg atctacaaag atgctcttgt tctacacaaa gtcctgcttg aaacacgcag
     3361 agacctggag ggagatgagg actctcatgt cccaaatgtg actttgctga ttcaagagct
     3421 tatccacaat ctttttgtgt cagtcatgag tcatcaggat gatgagggaa gatgctacag
     3481 cgattcttta gcagaaattc ctgctgtgga tcccaacttt cctaacaaac caccccttac
     3541 atttgacata attaggaaga atgttgaaaa taatcgctac cgtcggcttg atttatttca
     3601 agagcatatg tttgaagtat tggaacgagc aagaaggatg aatcggacag attcagaaat
     3661 atatgaagat gcagtagaac ttcagcagtt ttttattaaa attcgtgatg aactctgcaa
     3721 aaatggagag attcttcttt caccggcact cagctatacc acaaaacatt tgcataatga
     3781 tgtggagaaa gagagaaagg aaaaattgcc aaaagaaata gaggaagata aactaaaacg
     3841 agaagaagaa aaaagagaag ctgaaaagag tgaagattcc tctggtgctg caggcctctc
     3901 aggcttacat cgcacataca gccaggactg tagctttaaa aacagcatgt accatgttgg
     3961 agattacgtc tatgtggaac ctgcagaggc caacctacaa ccacatatcg tctgtattga
     4021 aagactgtgg gaggattcag ctggtgaaaa atggttgtat ggctgttggt tttaccgacc
     4081 aaatgaaaca ttccacctgg ctacacgaaa atttctagaa aaagaagttt ttaagagtga
     4141 ctattacaac aaagttccag ttagtaaaat tctaggcaag tgtgtggtca tgtttgtcaa
     4201 ggaatacttt aagttatgcc cagaaaactt ccgagatgag gatgtttttg tctgtgaatc
     4261 acggtattct gccaaaacca aatcttttaa gaaaattaaa ctgtggacca tgcccatcag
     4321 ctcagtcagg tttgtccctc gggatgtgcc tctgcctgtg gttcgcgtgg cctctgtatt
     4381 tgcaaatgca gataaaggtg atgatgagaa gaatacagac aactcagagg acagtcgagc
     4441 tgaagacaat tttaacttgg aaaaggaaaa agaagatgtc cctgtggaaa tgtccaatgg
     4501 tgaaccaggt tgccactact ttgagcagct ccattacaat gacatgtggc tgaaggttgg
     4561 cgactgtgtc ttcatcaagt cccatggcct ggtgcgtcct cgtgtgggca gaattgaaaa
     4621 agtatgggtt cgagatggag ctgcatattt ttatggcccc atcttcattc accctgaaga
     4681 aacagagcat gagcccacaa aaatgttcta caaaaaagaa gtatttctga gtaatctgga
     4741 agaaacctgc cccatgacat gtattctcgg aaagtgtgct gtgttgtcat tcaaggactt
     4801 cctctcctgc aggccaactg aaataccaga aaatgacatt ctgctttgtg agagccgcta
     4861 caatgagagc gacaagcaga tgaagaaatt caaaggattg aagaggtttt cactctctgc
     4921 taaagtggta gatgatgaaa tttactactt cagaaaacca attgttcctc agaaggagcc
     4981 atcacctttg ctggaaaaga agatccagtt gctagaagct aaatttgccg agttagaagg
     5041 tggagatgat gatattgaag agatgggaga agaagatagt gaggtcattg aacctccttc
     5101 tctacctcag cttcagaccc ccctggccag tgagctggac ctcatgccct acacaccccc
     5161 acagtctacc ccaaagtctg ccaaaggcag tgcaaagaag gaaggctcca aacggaaaat
     5221 caacatgagt ggctacatcc tgttcagcag tgagatgagg gctgtgatta aggcccaaca
     5281 cccagactac tctttcgggg agctcagccg cctggtgggg acagaatgga gaaatcttga
     5341 gacagccaag aaagcagaat atgaaggcat gatgggtggc tatccgccag gccttccacc
     5401 tttgcagggc ccagttgatg gccttgttag catgggcagc atgcagccac ttcaccctgg
     5461 ggggcctcca ccccaccatc ttccgccagg tgtgcctggc ctcccgggca tcccaccacc
     5521 gggtgtgatg aaccaaggag tggcccctat ggtagggact ccagcaccgg gtggaagtcc
     5581 atatggacaa caggtgggag ttttggggcc tccagggcag caggcaccac ctccatatcc
     5641 cggcccacat ccagctggac cccctgtcat acagcagcca acaacaccca tgtttgtagc
     5701 tcccccacca aagacccagc ggcttcttca ctcagaggcc tacctgaaat acattgaagg
     5761 actcagtgcg gagtccaaca gcattagcaa gtgggatcag acactggcag ctcgaagacg
     5821 cgacgtccat ttgtcgaaag aacaggagag ccgcctaccc tctcactggc tgaaaagcaa
     5881 aggggcccac accaccatgg cagatgccct ctggcgcctt cgagatttga tgctccggga
     5941 caccctcaac attcgccaag catacaacct agaaaatgtt taagcggccg cagaattcgc
     6001 ggccgcactc gagatatcta gacccagctt tcttgtacaa agtggttgat atccagcaca
     6061 gtggcggccg ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     6121 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6181 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6241 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6301 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6361 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6421 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6481 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6541 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6601 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6661 cccgcctgga attctgcaga tatccggact agtgatctct cgaggttaac gaattctacc
     6721 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     6781 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     6841 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     6901 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     6961 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     7021 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     7081 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     7141 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     7201 cctcatctcc gggcctttcg acctgcatcc cgccaccatg aaaaagcctg aactcaccgc
     7261 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     7321 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     7381 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     7441 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     7501 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     7561 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     7621 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     7681 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     7741 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     7801 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     7861 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     7921 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     7981 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     8041 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     8101 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     8161 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     8221 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa
     8281 caagagctga tttcgagaac gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg
     8341 cgagagaacg gccttacgct tggtggcaca gttctcgtcc acagttcgct aagctcgctc
     8401 ggctgggtcg cgggagggcc ggtcgcagtg attcaggccc ttctggattg tgttggtccc
     8461 cagggcacga ttgtcatgcc cacgcactcg ggtgatctga ctgatcccgc agattggaga
     8521 tcgccgcccg tgcctgccga ttgggtgcag atccgtcgag ggcccggtac ctttaagacc
     8581 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     8641 agggctagct cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     8701 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     8761 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     8821 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     8881 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     8941 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     9001 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     9061 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     9121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     9181 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     9241 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     9301 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     9361 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     9421 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     9481 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     9541 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     9601 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     9661 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     9721 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     9781 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     9841 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     9901 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     9961 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    10021 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
    10081 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
    10141 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
    10201 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
    10261 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
    10321 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
    10381 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
    10441 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
    10501 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
    10561 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
    10621 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    10681 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    10741 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    10801 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    10861 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    10921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    10981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    11041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    11101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    11161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
    11221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    11281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    11341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    11401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
    11461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
    11521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
    11581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
    11641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
    11701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
    11761 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
    11821 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
    11881 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
    11941 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
    12001 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
    12061 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
    12121 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
    12181 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
    12241 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
    12301 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
    12361 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
    12421 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
    12481 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
    12541 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
    12601 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
    12661 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
    12721 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
    12781 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
    12841 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
    12901 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
    12961 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
    13021 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
    13081 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
    13141 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
    13201 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
    13261 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
    13321 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
    13381 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
    13441 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
    13501 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
    13561 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
    13621 gaattattgg aattagataa atgggcaagt ttgtgg
//
