LOCUS       sequence-384389-       11221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR-assisted HDR, Rosa26 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11221)
  AUTHORS   Freed-Pastor WA, Lambert LJ, Ely ZA, Pattada NB, Bhutkar A, Eng G, 
            Mercer KL, Garcia AP, Lin L, Rideout WM 3rd, Hwang WL, Schenkel JM, 
            Jaeger AM, Bronson RT, Westcott PMK, Hether TD, Divakar P, Reeves 
            JW, Deshpande V, Delorey T, Phillips D, Yilmaz OH, Regev A, Jacks T
  TITLE     The CD155/TIGIT axis promotes and maintains immune evasion in 
            neoantigen-expressing pancreatic cancer.
  JOURNAL   Cancer Cell. 2021 Oct 11;39(10):1342-1360.e14. doi: 
            10.1016/j.ccell.2021.07.007. Epub 2021 Aug 5.
  PUBMED    34358448
REFERENCE   2  (bases 1 to 11221)
  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: 
            "10.1016/j.ccell.2021.07.007"; journalName: "Cancer Cell"; date: 
            "2021-10-11- 11"; volume: "39"; issue: "10"; pages: "1342-1360.e14"
COMMENT     Sequence Label: U6-sgR26-eCas9-T2A-BlastR
FEATURES             Location/Qualifiers
     source          1..11221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             836..1016
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1063..1188
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2655..2903
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2655..2675
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2826..2845
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2924..2999
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3079..3290
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3322..3331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3331..3396
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3403..3423
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3448..5985,5986..5988,5989..6450,6451..6453,6454..6621,
                     6622..6624,6625..7548)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7549..7596
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7606..7659
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7660..8058
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8111..8699
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8164..8184)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8582..8593)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8702..8718)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8703..8719)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9228..9408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(9470..10058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9550..9569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10229..11020,11021..11089))
                     /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     10852..10871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11090..11194)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
      841 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      901 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      961 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     1021 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     1081 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1141 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1201 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1261 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     1321 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1381 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1441 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1501 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc ccgggtcgac tagtgatcag tgtgagggag
     2641 tgtaaagctg gtttgagggc ctatttccca tgattccttc atatttgcat atacgataca
     2701 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     2761 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     2821 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     2881 tcttgtggaa aggacgaaac accggattct cccaggccca ggggttttag agctagaaat
     2941 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     3001 tttttaaacc gctgttccta ggaatcccga ggccttaggt cttgaaagga gtgggaattg
     3061 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     3121 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     3181 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     3241 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ggacccgaca
     3301 ttagcgctac agcttaagcg ggccaccatg gactataagg accacgacgg agactacaag
     3361 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
     3421 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     3481 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     3541 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     3601 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     3661 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     3721 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     3781 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     3841 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     3901 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     3961 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     4021 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     4081 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     4141 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     4201 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     4261 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     4321 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     4381 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     4441 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     4501 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     4561 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     4621 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     4681 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     4741 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     4801 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     4861 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     4921 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     4981 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     5041 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     5101 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     5161 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     5221 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     5281 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     5341 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     5401 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     5461 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     5521 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     5581 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     5641 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     5701 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     5761 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     5821 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     5881 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     5941 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctggccga cgactccatc
     6001 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     6061 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     6121 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     6181 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     6241 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     6301 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     6361 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     6421 gtgggaaccg ccctgatcaa aaagtaccct gcgctggaaa gcgagttcgt gtacggcgac
     6481 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     6541 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     6601 gccaacggcg agatccggaa ggcgcctctg atcgagacaa acggcgaaac cggggagatc
     6661 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     6721 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     6781 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     6841 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     6901 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     6961 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     7021 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     7081 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     7141 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     7201 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     7261 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     7321 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     7381 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     7441 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     7501 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     7561 gccacgaaaa aggccggcca ggcaaaaaag aaaaagggca gtggagaggg cagaggaagt
     7621 ctgctaacat gcggtgacgt cgaggagaat cctggcccaa tggccaagcc tttgtctcaa
     7681 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     7741 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     7801 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     7861 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     7921 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     7981 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     8041 tatgtgtggg agggctaaaa acgttgttgt ttggggttga attactcttc tagaccgcgg
     8101 cgcgcccgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     8161 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     8221 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     8281 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     8341 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     8401 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     8461 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     8521 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     8581 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     8641 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     8701 tcgataccgt cgacctcgat cgagacctag aaaaacatgg agcaatcaca agtagcaata
     8761 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     8821 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     8881 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     8941 atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca
     9001 caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     9061 ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg
     9121 tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc
     9181 gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt
     9241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     9301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     9361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa
     9421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     9481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     9541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     9601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     9661 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     9721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     9781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     9841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     9901 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     9961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
    10021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
    10081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
    10141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
    10201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
    10261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
    10321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
    10381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
    10441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
    10501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
    10561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
    10621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
    10681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
    10741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
    10801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
    10861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
    10921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    10981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    11041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
    11101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
    11161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
    11221 a
//
