LOCUS       sequence-314330-        9889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine Fbxw7 alpha with a T199 silent mutation in 
            mammalian cells. The T199 silent mutation confers resistance to the 
            5'-TGAACATGGTACAAGGCCAG-3' sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9889)
  AUTHORS   Gstalder C, Liu D, Miao D, Lutterbach B, DeVine AL, Lin C, Shettigar
            M, Pancholi P, Buchbinder EI, Carter SL, Manos MP, Rojas-Rudilla V, 
            Brennick R, Gjini E, Chen PH, Lako A, Rodig S, Yoon CH, Freeman GJ, 
            Barbie DA, Hodi FS, Miles W, Van Allen EM, Haq R
  TITLE     Inactivation of Fbxw7 impairs dsRNA sensing and confers resistance 
            to PD-1 blockade.
  JOURNAL   Cancer Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416.
  PUBMED    32371478
REFERENCE   2  (bases 1 to 9889)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer 
            Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416."
COMMENT     Sequence Label: pLX304_zeo_mmFbxw7(res)
FEATURES             Location/Qualifiers
     source          1..9889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..501
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     420..438
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             513..887
                     /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"
     misc_feature    947..1064
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1181..1484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1485..1688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1639..1659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1711..1735
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3924..3948)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3950..3991
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4033..4621
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4086..4106)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4504..4515)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4693..4926
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5004..5125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5041..5060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5095..5114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5165..5300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5320..5339)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5321..5339)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5349..5366)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5349..5365)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5358..5380)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5507..5962
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5594..5613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5804..5825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5988..6092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6093..6161,6162..6953)
                     /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(6311..6330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7124..7712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7613..7632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7866..7883
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8000..8021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8036..8041,8042..8059,8060..8066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8074..8090
                     /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     8079..8101
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8098..8114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8098..8114
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8133..8153
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8135..8153
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8181..8407
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8408..8588
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8635..8760
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9253..9486
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9671..9715
                     /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 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
       61 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
      121 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
      181 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
      241 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
      301 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
      361 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
      421 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
      481 atcaccgacc tctctcccca gggggtacca ccatggccaa gttgaccagt gccgttccgg
      541 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
      601 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
      661 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
      721 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
      781 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
      841 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgactg cagctgcagt
      901 agtaagaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      961 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1021 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     1081 acagcagaga tccactttgg cgccggctcg agggggcccg gggttattaa tagtaatcaa
     1141 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1201 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1261 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1321 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1381 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1441 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     1501 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     1561 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1621 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1681 gcagagctct ctggctaact gtcgggatca acaagtttgt acaaaaaagc aggctttaaa
     1741 ggaaccaatt cagtcgactg gatccggtac cgaattcgcg gccgcatgaa tcaggaactg
     1801 ctctctgtgg gcagcaaaag acgacgaact ggaggctctc tgagagggaa tgcttcctca
     1861 agccaggttg atgagggaca gatgaatcgc gtggttgagg aggatccaca gcagcaagcg
     1921 agacatcaag aggaggagca cactgcgcgg aatggtgaac ttgtgggtgc aaaccctagg
     1981 cctggagacc agaacgatac ccagcaagga caagtggagg aaaataataa ccgctttatt
     2041 tcagtagatg aggactcttc gggaaatcag gaagagcaag aggaagatga agagcatgct
     2101 ggggaacagg aggaggaaga ggaggaagag gaagaggagg aggagatgga ccaggagagt
     2161 gatgattttg atccgtctga tgacagtagc agagaagatg aacatacgca caatagcaat
     2221 gtcacaaact gcagtagtgt ctcggacctg cccgctcacc agctctcctc tccattctat
     2281 acaaagacaa caaaaatgaa aagaaagttg gaccatggtt ctgaagttcg ttccttttct
     2341 ttgggaaaga aaccatgcaa agtctcagat tataccagta caactggcct tgtaccatgt
     2401 tcagcaacac caacaacttt tggggacctg agagcagcca atgggcaagg gcagcagcgg
     2461 cggaggatta catctgtcca accacccaca ggccttcaag agtggctgaa aatgtttcag
     2521 agctggagcg gaccagagaa gttgctggct ttagatgagc tcattgacag ctgtgaacca
     2581 acacaagtga agcatatgat gcaagtgata gagccccagt tccagcgaga cttcatctcc
     2641 ttgcttccta aagagttggc actctatgtg ctttcattcc tggaacccaa agacctgctg
     2701 caagcggctc agacttgtcg atactggaga attttggctg aggataacct tctctggaga
     2761 gagaaatgta aagaagaggg gattgatgaa ccgttgcaca tcaagagaag aaaaataata
     2821 aaaccaggtt tcatacacag cccatggaag agtgcgtata tcagacagca cagaattgat
     2881 acaaactgga gacgaggaga actcaaatct cctaaggtgc tgaaagggca tgatgaccat
     2941 gtgatcacat gcctacagtt ttgtggcaac cgcatagtta gtggttctga tgacaacact
     3001 ttaaaagttt ggtcagcggt cacgggcaag tgtctgagaa cgttagtggg acatacaggt
     3061 ggagtgtggt catcacagat gagagacaat atcatcatca gtggatcgac tgaccggact
     3121 ctcaaagtgt ggaatgctga aactggagag tgtatacata ctttatatgg gcacacttct
     3181 actgtacggt gtatgcatct ccatgaaaaa agggttgtaa gcggttctcg agatgccact
     3241 ctcagggttt gggatattga gaccggccag tgtttacacg tcctgatggg tcacgtagca
     3301 gcggtccgct gcgttcagta tgatggcagg agggttgtta gtggagctta tgattttatg
     3361 gtgaaggtgt gggatccaga gactgagacc tgtctacaca cgttacaggg acacactaat
     3421 agagtctatt cattacagtt tgatggcatc catgtggtga gtggatctct tgatacatca
     3481 atccgagtct gggatgtgga gacagggaat tgtattcaca cgctaacagg acaccagtca
     3541 ttaacgagtg gaatggaact caaagacaat attcttgtct ctgggaatgc agattctaca
     3601 gttaagatct gggatatcaa aacaggacag tgtttacaaa ctttgcaagg tcccagcaag
     3661 catcagagcg ctgtgacctg cttacagttc aacaagaact tcgtaattac cagctcagac
     3721 gacggaacgg tcaaactctg ggacttgaaa acgggtgaat ttatccgaaa cctcgtcaca
     3781 ttggagagtg gggggagcgg gggagttgtg tggcggatca gggcctcaaa cacaaagctg
     3841 gtgtgtgcag tcgggagtcg gaatggaact gaggaaacca agctcctggt gctggacttt
     3901 gatgtggaca tgaaatgatc tagacccagc tttcttgtac aaagtggttg gtaagcctat
     3961 ccctaaccct ctcctcggtc tcgattctac gtagtaatga gctagcgcta accggtggcg
     4021 cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4081 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4141 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4201 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4261 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4321 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4381 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4441 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4501 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4561 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4621 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     4681 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     4741 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     4801 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4861 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4921 ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     4981 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     5041 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5101 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     5161 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5221 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5281 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     5341 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5401 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5461 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     5521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5701 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5761 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5821 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5881 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5941 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6001 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6061 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6121 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6181 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6241 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6301 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6361 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6421 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6481 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6541 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6601 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6661 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6721 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6781 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6841 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6901 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6961 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7021 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7081 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7141 tgcgcataat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7201 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7261 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7321 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7381 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7441 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7501 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7561 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7621 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7681 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7741 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7801 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7861 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7921 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7981 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     8041 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     8101 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     8161 agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt
     8221 aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt
     8281 ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat
     8341 tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata
     8401 cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     8461 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     8521 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     8581 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     8641 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     8701 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     8761 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     8821 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     8881 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     8941 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     9001 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     9061 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     9121 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     9181 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     9241 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     9301 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     9361 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     9421 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     9481 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     9541 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     9601 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     9661 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     9721 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     9781 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     9841 atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccc
//
