LOCUS       sequence-318003-       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        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..2172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2179..2199
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2251..2436
                     /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(2458..2481,2482..2484,2485..5040,5041..5043,
                     5044..6558)
                     /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             6559..6606
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6622..6678
                     /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             6679..7389
                     /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(6824..6842)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7092..7111
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    7406..7994
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7459..7479)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7877..7888)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8066..8299
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8377..8498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8414..8433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8468..8487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8538..8673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8693..8712)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8694..8712)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8722..8739)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8722..8738)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8731..8753)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8880..9335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8967..8986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9177..9198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9361..9465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9466..9534,9535..10326)
                     /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(9684..9703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10497..11085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10986..11005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11239..11256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11373..11394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11409..11414,11415..11432,11433..11439)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11447..11463
                     /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     11452..11474
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11471..11487
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11471..11487
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11506..11526
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11508..11526
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ttacaaagac
     2161 gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca
     2221 gccgatgcta agtcactaac tgcctggtcc cggacactgg tgaccttcaa ggatgtattt
     2281 gtggacttca ccagggagga gtggaagctg ctggacactg ctcagcagat cgtgtacaga
     2341 aatgtgatgc tggagaacta taagaacctg gtttccttgg gttatcagct tactaagcca
     2401 gatgtgatcc tccggttgga gaagggagaa gagcccggcg gttccggcgg agggtcggac
     2461 aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc
     2521 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     2581 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     2641 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     2701 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     2761 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     2821 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     2881 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     2941 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     3001 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     3061 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     3121 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt
     3181 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     3241 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     3301 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     3361 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     3421 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     3481 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     3541 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     3601 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     3661 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgcaatt
     3721 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     3781 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     3841 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     3901 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     3961 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     4021 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     4081 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     4141 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     4201 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     4261 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     4321 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     4381 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     4441 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     4501 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     4561 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     4621 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     4681 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     4741 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     4801 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     4861 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     4921 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccacatcgtg
     4981 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     5041 gcccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     5101 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     5161 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     5221 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     5281 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     5341 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     5401 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     5461 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     5521 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     5581 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     5641 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     5701 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     5761 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     5821 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     5881 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     5941 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     6001 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     6061 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     6121 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     6181 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     6241 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     6301 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     6361 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     6421 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     6481 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     6541 tctcagctgg gaggcgacaa gcgtcctgct gctactaaga aagctggtca agctaagaaa
     6601 aagaaagcta gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg
     6661 gaggagaacc ccggccccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     6721 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     6781 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     6841 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     6901 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     6961 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     7021 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     7081 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     7141 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     7201 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     7261 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     7321 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     7381 tacaagtaag acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     7441 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     7501 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     7561 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     7621 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     7681 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     7741 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     7801 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     7861 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     7921 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     7981 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     8041 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     8101 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     8161 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     8221 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     8281 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     8341 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     8401 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     8461 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     8521 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     8581 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     8641 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     8701 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     8761 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     8821 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     8881 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     8941 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     9001 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     9061 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     9121 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     9181 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     9241 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     9301 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     9361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     9421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     9481 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     9541 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     9601 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     9661 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     9721 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     9781 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     9841 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     9901 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     9961 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    10021 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    10081 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    10141 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    10201 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    10261 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    10321 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    10381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    10441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    10501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    10561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    10621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    10681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    10741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    10801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    10861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    10921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    10981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    11041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    11101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
    11161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
    11221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
    11281 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
    11341 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
    11401 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
    11461 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
    11521 taaagggaac aaaagctgga gctgcaagc
//
