LOCUS       sequence-474409-       10771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal fusion of HaloTag to dSpCas9, mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10771)
  TITLE     A Split Luciferase Biosensing Platform for Detection and Imaging of 
            Chromatin Loops in Individual Live Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10771)
  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: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-dSpCas9
FEATURES             Location/Qualifiers
     source          1..10771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1337..2131)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1448..1467)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2058..2077
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2198..2527)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2212..2347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2266..2285)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2512..2532
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2541..2969)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2651..2672)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2863..2882
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3171..3219
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3244..3263
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3327..3383)
                     /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             complement(3402..3449)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(3525..5039,5040..5042,5043..5108,5109..5114,
                     5115..7598,7599..7601,7602..7628))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGDTAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3525..7628)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGDTAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7689..8579)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             complement(8604..8630)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(8649..8669)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8766..8790)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(8787..8990)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8816..8836)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        8991..9370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9542..9561
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9636..9740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9741..9809,9810..10601)
                     /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(9959..9978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgcg
     1321 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     1381 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     1441 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     1501 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     1561 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     1621 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     1681 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     1741 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     1801 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     1861 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     1921 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     1981 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     2041 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     2101 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     2161 cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa
     2221 aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg gcctctgcat
     2281 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     2341 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     2401 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     2461 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     2521 gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2581 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2641 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2701 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2761 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2821 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2881 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2941 cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc agctgcgcag
     3001 atctgctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc cttcacccga
     3061 acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc aatggggtct
     3121 cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga
     3181 cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     3241 gtattgtctc cttccgtgtt tcagttagcc tcccccgttt aaactcatta ctaaccggta
     3301 gggatcgaac cctttcatgc ggccgcaggt ccagggttct cctccacgtc cccagcctgc
     3361 ttcagcaggc tgaagttagt agctccgctt ccactaccac cctttttctt ttttgcctga
     3421 ccggcctttt tcgtggccgc cggcctttta gatccacctc cggagcctcc accgctagcg
     3481 ctccctccgc tccccccgga tcctcctgaa cctccactac caccgtcccc accgagctga
     3541 gagaggtcga ttcttgtttc atagagcccc gtaattgact gatgaatcag tgtggcgtcc
     3601 aggacctcct ttgtagaggt gtaccgcttt ctgtctatgg tggtgtcgaa gtacttgaag
     3661 gctgcaggcg cgcccaagtt ggtcagagta aacaagtgga taatgttttc tgcctgctcc
     3721 ctgatgggct tatccctgtg cttattgtaa gcagaaagca ccttatcgag gttagcgtcg
     3781 gcgaggatca ctcttttgga gaattcgctt atttgctcga tgatctcatc aaggtagtgt
     3841 ttgtgttgtt ccacgaacag ctgcttctgc tcattatctt cgggagaccc tttgagcttt
     3901 tcatagtggc tggccagata caagaaatta acgtatttag agggcagtgc cagctcgtta
     3961 cctttctgca gctcgcccgc actagcgagc attcgtttcc ggccgttttc aagctcaaag
     4021 agagagtact tgggaagctt aatgatgagg tcttttttga cctctttata tcctttcgcc
     4081 tcgagaaagt cgatggggtt tttttcgaag cttgatcgct ccatgattgt gatgcccagc
     4141 agttccttga cgcttttgag ttttttagac ttccctttct ccactttggc cacaaccagt
     4201 acactgtaag cgactgtagg agaatcgaat ccgccgtatt tcttggggtc ccaatctttt
     4261 ttgcgtgcga tcagcttgtc gctgttcctt ttcgggagga tactttcctt ggagaagcct
     4321 ccggtctgta cttcggtctt tttaacgatg ttcacctgcg gcatggacag gaccttccgg
     4381 actgtcgcga aatccctacc cttgtcccac acgatttctc ctgtttctcc gtttgtttcg
     4441 ataagtggtc gcttccgaat ctctccattg gccagtgtaa tctcggtctt gaaaaaattc
     4501 ataatattgc tgtaaaagaa gtacttagcg gtggccttgc ctatttcctg ctcagacttt
     4561 gcgatcattt tcctaacatc gtacacttta tagtctccgt aaacaaattc agattcaagc
     4621 ttgggatatt ttttgataag tgcagtgcct accactgcat tcaggtaggc atcatgcgca
     4681 tggtggtaat tgttgatctc tctcacctta taaaactgaa agtcctttct gaaatctgag
     4741 accagcttag acttcagagt aataactttc acctctcgaa tcagtttgtc attttcatcg
     4801 tacttggtgt tcatgcgtga atcgagaatt tgggccacgt gcttggtgat ctggcgtgtc
     4861 tcaacaagct gccttttgat gaagccggct ttatccaact cagacaggcc acctcgttca
     4921 gccttagtca gattatcgaa cttccgttgt gtgatcagtt tggcgttcag cagctgccgc
     4981 caataatttt tcattttctt gacaacttct tctgagggga cgttatcact cttccctcta
     5041 tttttatcgg atcttgtcaa cactttatta tcaatagaat catctttgag aaaagactgg
     5101 ggcacgatag catccacgtc gtagtcggag agccgattga tgtccagttc ctgatccacg
     5161 tacatgtccc tgccgttctg caggtagtac aggtagagct tctcattctg aagctgggtg
     5221 ttttcaactg ggtgttcctt aaggatttgg gaccccagtt cttttatacc ctcttcaatc
     5281 ctcttcatcc tttccctact gttcttctgt cccttctggg tagtttggtt ctctcgggcc
     5341 atctcgataa cgatattctc gggcttatgc cttcccatta ctttgacgag ttcatccacg
     5401 accttaacgg tctgcagtat tccctttttg atagctgggc tacctgcaag attagcgatg
     5461 tgctcgtgaa gactgtcccc ctggccagaa acttgtgctt tctggatgtc ctccttaaag
     5521 gtgagagagt catcatggat caactgcatg aagttcctgt tggcaaatcc atcggactta
     5581 agaaaatcca ggattgtctt tccactctgc ttgtctcgga tcccattgat cagttttctt
     5641 gacagccgcc cccatcctgt atatcggcgc ctcttgagct gtttcatgac tttgtcgtcg
     5701 aagagatgag cgtaagtttt caagcgttct tcaatcatct ccctatcttc aaacaacgta
     5761 agggtgagga caatgtcctc aagaatgtcc tcgttctcct cattgtccag gaagtccttg
     5821 tctttaatga ttttcaggag atcgtgatac gttcccaggg atgcgttgaa gcgatcctcc
     5881 actccgctga tttcaacaga gtcgaaacat tcaatctttt tgaaatagtc ttctttgagc
     5941 tgtttcacgg taactttccg gttcgtcttg aagaggaggt ccacgatagc tttcttctgc
     6001 tctccagaca ggaatgctgg ctttctcatc ccttctgtga cgtatttgac cttggtgagc
     6061 tcgttataaa ctgtgaagta ctcgtacagc agagagtgtt taggaagcac cttttcgtta
     6121 ggcagatttt tatcaaagtt agtcatcctt tcgatgaagg actgggcaga ggccccctta
     6181 tccacgactt cctcgaagtt ccagggagtg atggtctctt ctgatttgcg agtcatccac
     6241 gcgaatctgg aatttccccg ggcgaggggg cctacatagt agggtatccg aaatgtgagg
     6301 attttctcaa tcttttccct gttatctttc aaaaaggggt agaaatcctc ttgccgcctg
     6361 aggatagcgt gcagttcgcc caggtgaatc tggtggggga tgcttccatt gtcgaaagtg
     6421 cgctgtttgc gcaacagatc ttctctgtta agctttacca gcagctcctc ggtgccgtcc
     6481 attttttcca agatgggctt aataaatttg taaaattcct cctggcttgc tccgccgtca
     6541 atgtatccgg cgtagccatt tttagactga tcgaagaaaa tttccttgta cttctcaggc
     6601 agttgctgtc tgacaagggc cttcagcaaa gtcaagtctt ggtggtgctc atcatagcgc
     6661 ttgatcatac tagcgctcag cggagctttg gtgatctccg tgttcactcg cagaatatca
     6721 ctcagcagaa tggcgtctga caggttcttt gccgccaaaa aaaggtctgc gtactggtcg
     6781 ccgatctggg ccagcagatt gtcgagatca tcatcgtagg tgtctttgct cagttgaagc
     6841 ttggcatctt cggccaggtc gaagttagat ttaaagttgg gggtcagccc gagtgacagg
     6901 gcgataagat taccaaacag gccgttcttc ttctccccag ggagctgtgc gatgaggttt
     6961 tcgagccgcc gggatttgga cagcctagcg ctcaggattg ctttggcgtc aactccggat
     7021 gcgttgatcg ggttctcttc gaaaagctga ttgtaagtct gaaccagttg gataaagagt
     7081 ttgtcgacat cgctgttgtc tgggttcagg tccccctcga tgaggaagtg tccccgaaat
     7141 ttgatcatat gcgccagcgc gagatagatc aaccgcaagt cagccttatc agtactgtct
     7201 acaagcttct tcctcagatg atatatggtt gggtactttt catggtacgc cacctcgtcc
     7261 acgatattgc caaagattgg gtggcgctcg tgctttttat cctcctccac caaaaaggac
     7321 tcctccagcc tatggaagaa agagtcatcc accttagcca tctcattact aaagatctcc
     7381 tgcaggtagc agatccgatt ctttctgcgg gtatatctgc gccgtgctgt tcttttgagc
     7441 cgcgtggctt cggccgtgtc cccggagtcg aacaggaggg cgccaatgag gttcttcttt
     7501 atgctgtggc gatcggtatt gcccagaact ttgaattttt tgctcggcac cttgtactcg
     7561 tccgtaatga cggcccagcc gacgctgttt gtgccgatag cgagcccaat ggagtacttc
     7621 ttgtccattg gccggccgct gccgcctgag ccaccagaac cgccgcttcc accactccct
     7681 ccggtaccgc cggaaatctc gagcgtcgac agccagcgcg cgatctcgct gccgatcagg
     7741 tccgggttgt cttcttgcag cagattcaga cccgggccga tgtccacagc cttgcagtta
     7801 ggcaggcttt tggccaggcg agcggcttcg gccggtggga tcagaacgcc tggggtgccc
     7861 cagaacagca gcttcgggac aggggactgg tgcagccagt ccatgtattc ttcgaccagc
     7921 gcgacgatgt tcgctggctc accggcgatt ggcagctcgt ttgggaagcg ccacagtggc
     7981 tcgcggtcaa caggattcag gaacggctcg cggtaatggt ccatctcgac ttcagtcagc
     8041 gggcggacga cacccatcgg cagcgtaccc tcgataaaaa cgttctgatc gatgatcagc
     8101 ttgcggccga cgtcggtggt gcggaaggcc tggaaggtct cgcgggcaaa ttctggccat
     8161 tcgtcccagg tcgggatagg gcggatgaac tccataaatg caataccttt gacgcgctct
     8221 ggattgcgct tggcccagtg gaaacccaga gcggagcccc agtcgtgaat gaccaggacg
     8281 acctcttcca gacccagggc ttcgatgaag gcatccatga agcggacgtg gtcgtcgaag
     8341 aaataaccca ggtctggttt gtcggatttg cccataccga tcaggtctgg agcaatgcag
     8401 cgatgggtcg gtgcaacatg cgggatgatg ttgcgccaca cgtaggagga ggtcgggtta
     8461 ccgtgcagga acagcacagg ggtgccatcg cgcggaccaa catcgacgta gtgcatgcgc
     8521 tcgcccagga cttccacata atgggggtcg aatggaaagc cagtaccgat ttctgccatg
     8581 gatccaccgc cggacccgcc acccgcgtag tctgggacat cgtatgggta gcttcctccg
     8641 gagcctccca ccttcctctt cttcttgggc atggtggcaa gggttcgatc ctctagagtc
     8701 cggaggctgg atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg atggcgtgtc
     8761 caggcgatct gacggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc
     8821 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     8881 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     8941 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     9001 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     9061 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     9121 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     9181 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     9241 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     9301 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     9361 aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc
     9421 ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg
     9481 ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat
     9541 gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc
     9601 cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     9661 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     9721 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     9781 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     9841 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     9901 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     9961 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
    10021 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
    10081 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
    10141 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
    10201 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
    10261 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
    10321 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    10381 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    10441 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
    10501 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    10561 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    10621 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    10681 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    10741 cagaccccgt agaaaagatc aaaggatctt c
//
