LOCUS       sequence-223274-       13500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-mediated activation of TCF7L2. Catalytically inactive Cas9 
            from S. pyogenes and VP64 with 2A-BlastR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13500)
  AUTHORS   Miguel-Escalada I, Bonas-Guarch S, Cebola I, Ponsa-Cobas J, 
            Mendieta-Esteban J, Atla G, Javierre BM, Rolando DMY, Farabella I, 
            Morgan CC, Garcia-Hurtado J, Beucher A, Moran I, Pasquali L, 
            Ramos-Rodriguez M, Appel EVR, Linneberg A, Gjesing AP, Witte DR, 
            Pedersen O, Grarup N, Ravassard P, Torrents D, Mercader JM, Piemonti
            L, Berney T, de Koning EJP, Kerr-Conte J, Pattou F, Fedko IO, Groop 
            L, Prokopenko I, Hansen T, Marti-Renom MA, Fraser P, Ferrer J
  TITLE     Human pancreatic islet three-dimensional chromatin architecture 
            provides insights into the genetics of type 2 diabetes.
  JOURNAL   Nat Genet. 2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 
            10.1038/s41588-019-0457-0.
  PUBMED    31253982
REFERENCE   2  (bases 1 to 13500)
  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: "Nat Genet. 
            2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 
            10.1038/s41588-019-0457-0."
COMMENT     Sequence Label: lentiSAMv2 TCF7L2 TSS-guide3
FEATURES             Location/Qualifiers
     source          1..13500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        18..36
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        88..106
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        193..404
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             421..468
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(469..492,493..495,496..3051,3052..3054,3055..4569)
                     /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             4624..4644
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4669..4818
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             4840..4893
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4894..5292
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5317..5905
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5370..5390)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5788..5799)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5908..5924)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5909..5925)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6430..6610
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6636..6653)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6642..6866
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6912..7340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6999..7018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7209..7230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7349..7369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7354..7683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7534..7669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7596..7615
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7731..7778
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7797..8171
                     /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    8301..8422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8338..8357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8392..8411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8471..8487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8471..8487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8484..8506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8495..8511
                     /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(8519..8525,8526..8543,8544..8549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8564..8585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8702..8719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8873..9461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8953..8972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9632..10423,10424..10492))
                     /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     10255..10274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10493..10597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10672..10691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10863..11242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11244..11442
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11397..11417
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11460..11640
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11687..11812
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12305..12538
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12723..12767
                     /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    13065..13182
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        13232..13480
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     13232..13252
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     13403..13422
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gttttagagc taggccaaca tgaggatcac ccatgtctgc agggcctagc aagttaaaat
       61 aaggctagtc cgttatcaac ttggccaaca tgaggatcac ccatgtctgc agggccaagt
      121 ggcaccgagt cggtgctttt tttggatcct aggtcttgaa aggagtggga attggctccg
      181 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
      241 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      301 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      361 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggcgtac ggccaccatg
      421 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga caagaagtac
      481 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
      541 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
      601 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
      661 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
      721 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
      781 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
      841 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
      901 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
      961 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     1021 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     1081 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     1141 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gcaacctgat tgccctgagc
     1201 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     1261 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     1321 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     1381 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     1441 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     1501 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     1561 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     1621 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     1681 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     1741 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     1801 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     1861 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     1921 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     1981 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     2041 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     2101 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     2161 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     2221 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     2281 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     2341 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     2401 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     2461 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     2521 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     2581 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     2641 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     2701 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     2761 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     2821 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     2881 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     2941 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accacatcgt gcctcagagc
     3001 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa ggcccggggc
     3061 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     3121 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     3181 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     3241 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     3301 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     3361 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     3421 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     3481 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     3541 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     3601 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     3661 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     3721 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     3781 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     3841 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     3901 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     3961 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     4021 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     4081 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     4141 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     4201 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     4261 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     4321 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     4381 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     4441 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     4501 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     4561 ggaggcgaca gcgctggagg aggtggaagc ggaggaggag gaagcggagg aggaggtagc
     4621 ggacctaaga aaaagaggaa ggtggcggcc gctggatccg gacgggctga cgcattggac
     4681 gattttgatc tggatatgct gggaagtgac gccctcgatg attttgacct tgacatgctt
     4741 ggttcggatg cccttgatga ctttgacctc gacatgctcg gcagtgacgc ccttgatgat
     4801 ttcgacctgg acatgctgat taactgtaca ggcagtggag agggcagagg aagtctgcta
     4861 acatgcggtg acgtcgagga gaatcctggc ccaatggcca agcctttgtc tcaagaagaa
     4921 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     4981 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     5041 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     5101 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     5161 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     5221 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     5281 tgggagggct aagaattcga tatcaagctt atcggtaatc aacctctgga ttacaaaatt
     5341 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5401 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5461 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5521 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5581 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5641 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5701 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5761 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5821 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5881 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat
     5941 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6001 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6061 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6121 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6181 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     6241 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     6301 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     6361 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     6421 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6481 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6541 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6601 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     6661 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     6721 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     6781 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     6841 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     6901 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     7021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7321 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     7381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     7441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     7561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     7621 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     7681 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     7741 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     7801 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     7861 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     7921 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     7981 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     8041 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     8101 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     8161 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     8221 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     8281 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     8341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8401 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     8461 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8521 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8581 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     8641 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8701 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8761 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8821 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     9001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     9061 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     9121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     9181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9301 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9361 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9421 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9481 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9541 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9601 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9661 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9721 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9781 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9841 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9901 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9961 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    10021 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    10081 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    10141 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    10201 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    10261 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10321 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10381 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10441 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10501 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10561 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    10621 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
    10681 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
    10741 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
    10801 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
    10861 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
    10921 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
    10981 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
    11041 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
    11101 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
    11161 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
    11221 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
    11281 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
    11341 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
    11401 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg
    11461 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
    11521 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
    11581 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
    11641 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
    11701 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
    11761 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
    11821 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
    11881 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
    11941 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
    12001 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
    12061 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
    12121 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
    12181 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
    12241 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
    12301 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
    12361 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
    12421 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
    12481 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
    12541 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
    12601 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
    12661 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
    12721 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
    12781 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
    12841 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
    12901 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
    12961 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
    13021 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
    13081 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
    13141 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
    13201 agcagagatc cagtttggtt aattagctag cgagggccta tttcccatga ttccttcata
    13261 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
    13321 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
    13381 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
    13441 tttcttggct ttatatatct tgtggaaagg acgaaacacc gcggaggaac ggagtagtct
//
