LOCUS       sequence-386206-       13713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tricistronic expression of dCas9-EGFP-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13713)
  AUTHORS   Teh SSK, Bowland K, Halper-Stromberg E, Kotwal A, Bennett A, Skaist 
            A, Tang J, Cai F, Macoretta A, Liang H, Kamiyama H, Wheelan S, Lin 
            MT, Hruban RH, Hung CF, Goldstein M, Scharpf RB, Roberts NJ, 
            Eshleman JR
  TITLE     CRISPR-Cas9 for selective targeting of somatic mutations in 
            pancreatic cancers.
  JOURNAL   NAR Cancer. 2024 Jun 19;6(2):zcae028. doi: 10.1093/narcan/zcae028. 
            eCollection 2024 Jun.
  PUBMED    38915758
REFERENCE   2  (bases 1 to 13713)
  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: "NAR 
            Cancer."; date: "2024-06-19"; volume: "6(2):zcae028. doi"; pages: " 
            10.1093/narcan/zcae028. eCollection 2024 Jun"
COMMENT     Sequence Label: pLenti-dCas9-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..13713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..396
                     /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"
     polyA_signal    526..647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(563..582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     617..636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(696..712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(696..712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(709..731)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    720..736
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(744..750,751..768,769..774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    789..810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(927..944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1098..1686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1178..1197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1857..2648,2649..2717))
                     /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     2480..2499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2718..2822)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2897..2916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3088..3467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3469..3667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3622..3642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3685..3865
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3912..4037
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4530..4763
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4948..4992
                     /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    5290..5407
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5500..5711
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5732..5797
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(5804..5827,5828..5830,5831..8317,8318..8320,
                     8321..9904)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5804..9904
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9905..9952
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9953..9976
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9986..10039
                     /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"
     regulatory      10046..10055
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(10052..10054,10055..10057,10058..10768)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKLSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(10358..10377)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10705..10726
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             10769..10786
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             10796..10852
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             10853..11251
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11276..11864
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11329..11349)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11747..11758)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11867..11883)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11868..11884)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12389..12569
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12595..12612)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12601..12825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12871..13299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12958..12977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13168..13189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13308..13328)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      13473..13607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13534..13553
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        join(13669..13713,1..3)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
       61 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
      121 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
      181 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
      241 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
      301 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
      361 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
      421 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
      481 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
      541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
      661 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      721 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      781 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2821 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     2881 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     2941 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     3001 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     3061 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     3121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     3301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     3361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     3421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     3481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     3541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     3601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     3661 taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
     3721 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     3781 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     3841 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     3901 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     3961 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     4021 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     4081 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     4141 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     4201 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     4261 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     4321 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     4381 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     4441 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     4501 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     4561 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     4621 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     4681 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     4741 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     4801 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     4861 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     4921 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     4981 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     5041 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     5101 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     5161 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     5221 attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt
     5281 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     5341 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     5401 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta gctagctagg
     5461 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     5521 cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc
     5581 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     5641 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     5701 ccagaacaca ggaccggtgc cgcccaccat ggactacaaa gaccatgacg gtgattataa
     5761 agatcatgac atcgattaca aggatgacga tgacaaggaa ttcgacaaga agtacagcat
     5821 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     5881 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     5941 gatcggcgcc ctgctgttcg acagcggaga aacagccgag gccacccggc tgaagagaac
     6001 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     6061 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     6121 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     6181 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     6241 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     6301 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     6361 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     6421 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     6481 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
     6541 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     6601 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     6661 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     6721 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     6781 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     6841 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacatcg atggcggagc
     6901 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     6961 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     7021 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     7081 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     7141 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     7201 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgccag
     7261 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     7321 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tacaacgagc tgaccaaagt
     7381 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaagc
     7441 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     7501 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     7561 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     7621 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     7681 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     7741 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     7801 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     7861 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     7921 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     7981 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     8041 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     8101 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     8161 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     8221 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     8281 actggacatc aaccggctgt ccgactacga tgtggacgct atcgtgcctc agagctttct
     8341 gaaggacgac tccatcgata acaaagtgct gactcggagc gacaagaacc ggggcaagag
     8401 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc gccagctgct
     8461 gaatgccaag ctgattaccc agaggaagtt cgacaatctg accaaggccg agagaggcgg
     8521 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     8581 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaacga
     8641 caaactgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     8701 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     8761 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     8821 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     8881 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     8941 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     9001 cgaaacaggc gagatcgtgt gggataaggg ccgggacttt gccaccgtgc ggaaagtgct
     9061 gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     9121 agagtctatc ctgcccaaga ggaacagcga caagctgatc gccagaaaga aggactggga
     9181 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     9241 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     9301 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     9361 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     9421 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     9481 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     9541 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaaac actacctgga
     9601 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     9661 ggacaaggtg ctgagcgcct acaacaagca cagagacaag cctatcagag agcaggccga
     9721 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     9781 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     9841 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     9901 cgacaagcga cctgccgcca caaagaaggc tggacaggct aagaagaaga aagattacaa
     9961 agacgatgac gataagggat ccggagaggg cagaggaagt ctgctaacat gcggtgacgt
    10021 cgaggagaat cctggcccac cggtcgccac catggtgagc aagggcgagg agctgttcac
    10081 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agctcagcgt
    10141 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
    10201 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
    10261 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
    10321 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
    10381 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
    10441 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
    10501 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
    10561 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
    10621 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
    10681 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
    10741 cactctcggc atggacgagc tgtacaagca tcatcaccat caccacggat ccggcgcaac
    10801 aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgatggccaa
    10861 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
    10921 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
    10981 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
    11041 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
    11101 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
    11161 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
    11221 gctgccctct ggttatgtgt gggagggcta agaattcgat atcaagctta tcggtaatca
    11281 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
    11341 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
    11401 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
    11461 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
    11521 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
    11581 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
    11641 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
    11701 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
    11761 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    11821 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
    11881 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
    11941 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
    12001 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    12061 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
    12121 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
    12181 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
    12241 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
    12301 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
    12361 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
    12421 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    12481 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    12541 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
    12601 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    12661 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    12721 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    12781 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    12841 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    12901 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    12961 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    13021 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    13081 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    13141 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    13201 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    13261 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    13321 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    13381 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    13441 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccagttccgc
    13501 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    13561 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
    13621 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
    13681 tcatcggcat agtatatcgg catagtataa tac
//
