LOCUS       sequence-324326-       11549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression plasmid of sgRNA with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11549)
  AUTHORS   Giles JR, Manne S, Freilich E, Oldridge DA, Baxter AE, George S, 
            Chen Z, Huang H, Chilukuri L, Carberry M, Giles L, Weng NP, Young 
            RM, June CH, Schuchter LM, Amaravadi RK, Xu X, Karakousis GC, 
            Mitchell TC, Huang AC, Shi J, Wherry EJ
  TITLE     Human epigenetic and transcriptional T cell differentiation atlas 
            for identifying functional T cell-specific enhancers.
  JOURNAL   Immunity. 2022 Mar 8;55(3):557-574.e7. doi: 
            10.1016/j.immuni.2022.02.004.
  PUBMED    35263570
REFERENCE   2  (bases 1 to 11549)
  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.immuni.2022.02"; journalName: "Immunity"; date: 
            "2022-03-8- 8"; volume: "55"; issue: "3"; pages: "557-574.e7"
COMMENT     Sequence Label: pED9x (dCas9-KRAB-mCherry)
FEATURES             Location/Qualifiers
     source          1..11549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      232..241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             241..264
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             271..291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             343..528
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             join(550..573,574..576,577..3132,3133..3135,3136..4650)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4651..4698
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4714..4770
                     /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             4771..5481
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4916..4934)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5184..5203
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5498..6086
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5551..5571)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5969..5980)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6158..6391
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6469..6590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6506..6525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6560..6579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6630..6765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6785..6804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6786..6804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6814..6831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6814..6830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6823..6845)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6972..7427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7059..7078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7269..7290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7453..7557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7558..7626,7627..8418)
                     /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(7776..7795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8589..9177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9078..9097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9331..9348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9465..9486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9501..9506,9507..9524,9525..9531)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9539..9555
                     /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     9544..9566
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9563..9579
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9563..9579
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9598..9618
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9600..9618
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9646..9872
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9873..10053
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10100..10225
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10718..10951
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11136..11180
                     /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    11364..11481
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
       61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      181 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg
      241 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
      301 ggagtcccag cagccgatgc taagtcacta actgcctggt cccggacact ggtgaccttc
      361 aaggatgtat ttgtggactt caccagggag gagtggaagc tgctggacac tgctcagcag
      421 atcgtgtaca gaaatgtgat gctggagaac tataagaacc tggtttcctt gggttatcag
      481 cttactaagc cagatgtgat cctccggttg gagaagggag aagagcccgg cggttccggc
      541 ggagggtcgg acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg
      601 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
      661 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
      721 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
      781 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
      841 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
      901 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
      961 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     1021 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     1081 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     1141 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     1201 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     1261 ggcaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     1321 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     1381 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     1441 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     1501 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     1561 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     1621 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     1681 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     1741 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     1801 ctgcacgcaa ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     1861 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     1921 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     1981 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     2041 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     2101 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     2161 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     2221 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     2281 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     2341 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     2401 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     2461 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     2521 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     2581 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     2641 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     2701 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     2761 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     2821 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     2881 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     2941 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     3001 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     3061 gaccacatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     3121 agaagcgaca aggcccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     3181 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     3241 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     3301 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     3361 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     3421 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     3481 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     3541 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     3601 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     3661 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     3721 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     3781 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     3841 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     3901 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     3961 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     4021 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     4081 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     4141 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     4201 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     4261 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     4321 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     4381 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     4441 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     4501 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     4561 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     4621 cggatcgacc tgtctcagct gggaggcgac aagcgtcctg ctgctactaa gaaagctggt
     4681 caagctaaga aaaagaaagc tagcggcagc ggcgccacca acttcagcct gctgaagcag
     4741 gccggcgacg tggaggagaa ccccggcccc atggtgagca agggcgagga ggataacatg
     4801 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     4861 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     4921 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     4981 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     5041 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     5101 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     5161 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     5221 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     5281 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     5341 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     5401 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     5461 atggacgagc tgtacaagta agacgcgtta agtcgacaat caacctctgg attacaaaat
     5521 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5581 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5641 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5701 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5761 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5821 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5881 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5941 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6001 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6061 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     6121 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6181 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6241 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6301 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6361 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt
     6421 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6601 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6661 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6721 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     6781 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     6841 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6901 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6961 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     7021 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     7081 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7141 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7201 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7261 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7321 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7381 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     7441 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     7501 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7561 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7621 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7681 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7741 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7801 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7861 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7921 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     7981 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8041 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8101 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8161 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8221 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     8281 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     8341 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     8401 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     8461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     8521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     8581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8701 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     8761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     8821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     8881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     8941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     9241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     9301 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     9361 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     9421 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     9481 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     9541 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     9601 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     9661 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     9721 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     9781 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     9841 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     9901 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     9961 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    10021 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
    10081 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
    10141 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
    10201 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
    10261 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
    10321 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
    10381 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
    10441 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
    10501 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
    10561 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
    10621 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
    10681 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
    10741 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
    10801 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
    10861 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
    10921 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
    10981 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
    11041 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
    11101 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
    11161 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
    11221 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
    11281 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
    11341 ccccgagggg acccagacca caattttaaa agaaaagggg ggattggggg gtacagtgca
    11401 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
    11461 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc
    11521 cggctcgagt ggctccggtg cccgtcagt
//
