LOCUS       sequence-190866-        9102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer/reporter plasmid for tissue-specific expression of Cas9 
            with 2A-Citrine reporter. Contains BsmBI-flanked LacZ cloning 
            cassette for rapid GoldenGate-based cloning of specific enhancers..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9102)
  AUTHORS   Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, 
            Sauka-Spengler T
  TITLE     Genome and epigenome engineering CRISPR toolkit for in vivo 
            modulation of cis-regulatory interactions and gene expression in the
            chicken embryo.
  JOURNAL   Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 
            10.1242/dev.160333.
  PUBMED    29386245
REFERENCE   2  (bases 1 to 9102)
  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: 
            "Development."; date: "2018-02-23"; volume: "145(4). pii: 
            dev.160333. doi"; pages: " 10.1242/dev.160333"
COMMENT     Sequence Label: pTK Cas9-2A-Citrine
FEATURES             Location/Qualifiers
     source          1..9102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    55..146
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     95..114
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     329..351
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     343..360
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     344..360
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     370..389
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        370..388
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(436..456)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(436..454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(475..491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(475..491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(488..510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    499..515
                     /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(523..529,530..547,548..553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    568..589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        757..902
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          980..1112
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1166..1175
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1178..1198
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1223..5323
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1223..5323
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5324..5371
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5387..5440
                     /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             join(5447..5449,5450..5452,5453..6163)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5753..5772)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6100..6121
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6218..6339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6229..6248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6283..6302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6587..6604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6758..7346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6838..6857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7517..8308,8309..8377))
                     /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     8140..8159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8378..8482)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8509..8964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8596..8615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8806..8827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(9042..9102,1..479))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLPGN
                     SSSVPIDREMFWHLHLHWGQPILLVCFVSFCFDGERMLVLSIHTKNQHTDVMKIKIFYC
                     GRSKYLPYLNITYLSW"
     polyA_signal    join(9095..9102,1..41)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
       61 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
      121 aagtgcaggt gccagaacat ttctctatcg ataggtaccg agctcgagtt gccagggaga
      181 cgggcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
      241 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
      301 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
      361 gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc
      421 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
      481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      601 ctcactgccc gctttccacc ggtcgtctcc tgttggctag cccgggctcg agatctgcga
      661 tctaagtaag cttgctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc
      721 ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc
      781 ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact
      841 tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt
      901 aacagcgtca acagcgtgcc gcagatccca agcttgcggc cgcttaagct gcagaagttg
      961 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     1021 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     1081 tactgacatc cactttgcct ttctctccac aggtgtccac tcccaggttc aatacagctc
     1141 ttaagcggcc gcaagctagc ttgccgccac catggcccca aagaagaagc ggaaggtcgg
     1201 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     1261 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     1321 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     1381 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     1441 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     1501 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     1561 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     1621 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     1681 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     1741 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     1801 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     1861 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     1921 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     1981 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     2041 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     2101 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     2161 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     2221 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     2281 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     2341 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     2401 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     2461 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     2521 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     2581 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     2641 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     2701 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     2761 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     2821 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     2881 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     2941 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     3001 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     3061 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     3121 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     3181 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     3241 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     3301 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     3361 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     3421 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     3481 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     3541 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     3601 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     3661 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     3721 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     3781 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     3841 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     3901 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     3961 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     4021 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     4081 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     4141 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     4201 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     4261 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     4321 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     4381 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     4441 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     4501 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     4561 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     4621 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     4681 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     4741 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     4801 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     4861 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     4921 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     4981 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     5041 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     5101 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     5161 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     5221 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     5281 cctgtacgag acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac
     5341 gaaaaaggcc ggccaggcaa aaaagaaaaa ggatatcgga tcaggagagg gcaggggaag
     5401 tcttctaaca tgcggggacg tggaggaaaa tcccgggccc agatccatgg tgagcaaggg
     5461 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     5521 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     5581 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     5641 cggctacggc ctgatgtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     5701 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5761 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     5821 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5881 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     5941 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     6001 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     6061 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     6121 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     6181 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     6241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     6301 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     6361 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     6421 aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt
     6481 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     6541 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
     6601 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6661 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6781 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8101 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8461 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     8521 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     8581 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     8641 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     8701 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     8761 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     8821 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     8881 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     8941 acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag
     9001 ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta
     9061 atattaaggt acgggaggta cttggagcgg ccgcaataaa at
//
