LOCUS       sequence-457355-        8505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Halo-tagged human BRD4 gRNA; CRISPR-mediated gene 
            insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8505)
  TITLE     Imielinski Lab CRISPR plasmids
REFERENCE   2  (bases 1 to 8505)
  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"
COMMENT     Sequence Label: pX330-U6-BRD4-HaloTag-Guide-hspCas9
FEATURES             Location/Qualifiers
     source          1..8505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..254
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6..26
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     177..196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        275..350
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        447..732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        734..1009
                     /label=chicken beta-actin promoter
     intron          1010..1237
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1249..1258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1258..1323
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1330..1350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1375..5475
                     /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             1375..5475
                     /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             5476..5523
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5551..5568)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5557..5764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5773..5913
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5773..5902
                     /label=AAV2 ITR
     rep_origin      5988..6443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6075..6094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6285..6306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6460..6479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6579..6601
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6639..6657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6725..6829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6830..6898,6899..7690)
                     /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(7048..7067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7861..8449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8350..8369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
       61 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      241 aaggacgaaa caccggggat cactagcatg tctggtttta gagctagaaa tagcaagtta
      301 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
      361 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
      421 tgcagacaaa tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat
      541 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      601 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt
      661 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      721 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
      781 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
      841 gggggggggg gggggggcgc ggggcggggc ggggcggggc gaggggcggg gcggggcgag
      901 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
      961 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
     1021 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
     1081 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
     1141 ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa
     1201 ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatggac
     1261 tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat
     1321 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
     1381 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     1441 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     1501 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     1561 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     1621 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     1681 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     1741 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     1801 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     1861 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1921 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     1981 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     2041 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     2101 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     2161 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     2221 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     2281 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     2341 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     2401 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     2461 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     2521 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     2581 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     2641 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     2701 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     2761 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     2821 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     2881 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     2941 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     3001 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     3061 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     3121 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     3181 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     3241 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     3301 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     3361 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     3421 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     3481 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     3541 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     3601 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     3661 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     3721 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     3781 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     3841 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     3901 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     3961 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     4021 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     4081 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     4141 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     4201 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     4261 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     4321 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     4381 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     4441 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     4501 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     4561 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     4621 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     4681 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     4741 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     4801 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     4861 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     4921 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     4981 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     5041 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     5101 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     5161 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     5221 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     5281 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     5341 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     5401 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     5461 cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa
     5521 aagtaagaat tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     5581 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5641 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5701 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     5761 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5881 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     5941 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     6001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6061 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6181 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     6241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     6301 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     6361 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     6421 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     6481 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6541 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6601 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     6661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     6721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6841 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6901 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6961 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7021 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7081 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7141 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7201 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7261 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7321 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7381 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7441 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7501 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     7561 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7621 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7681 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctca
//
