LOCUS       sequence-438226-        9646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Cas9-hGem and guideRNA targeting the human AAVS1 safe 
            harbour locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9646)
  AUTHORS   Batty P, Langer CC, Takacs Z, Tang W, Blaukopf C, Peters JM, Gerlich
            DW
  TITLE     Cohesin-mediated DNA loop extrusion resolves sister chromatids in G2
            phase.
  JOURNAL   EMBO J. 2023 Aug 15;42(16):e113475. doi: 10.15252/embj.2023113475. 
            Epub 2023 Jun 26.
  PUBMED    37357575
REFERENCE   2  (bases 1 to 9646)
  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: "EMBO J."; 
            date: "2023-08-15"; volume: "42(16):e113475. doi"; pages: " 
            10.15252/embj.2023113475. Epub 2023 Jun 26"
COMMENT     Sequence Label: AAVS1_Cas9-hGem_gRNA2
FEATURES             Location/Qualifiers
     source          1..9646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(517..537)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(544..609)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          630..857
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(858..1136)
                     /label=chicken beta-actin promoter
     enhancer        1138..1423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(1520..1595)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(1617..1865)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1675..1694)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1845..1865)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(1927..2515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2007..2026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2686..3477,3478..3546))
                     /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     3309..3328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3547..3651)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3719..3737
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3775..3797)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3897..3916
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3933..4388)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4070..4091)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4282..4301
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4463..4603
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4474..4603
                     /label=AAV2 ITR
     polyA_signal    4612..4819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4808..4825
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4850..5560)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     5397..5415
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5557..5566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5582..5638)
                     /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(5648..5977)
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     CDS             complement(5990..6037)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(6038..9646,1..492))
                     /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             complement(join(6038..9646,1..492))
                     /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"
ORIGIN
        1 ccggaacttg atcatgtggg ccagggccag atagatcagc cgcaggtcgg ccttgtcggt
       61 gctgtccacc agtttctttc tcaggtggta gatggtgggg tacttctcgt ggtaggccac
      121 ctcgtccacg atgttgccga agatggggtg ccgctcgtgc ttcttatcct cttccaccag
      181 gaaggactct tccagtctgt ggaagaagct gtcgtccacc ttggccatct cgttgctgaa
      241 gatctcttgc agatagcaga tccggttctt ccgtctggtg tatcttcttc tggcggttct
      301 cttcagccgg gtggcctcgg ctgtttcgcc gctgtcgaac agcagggctc cgatcaggtt
      361 cttcttgatg ctgtgccggt cggtgttgcc cagcaccttg aatttcttgc tgggcacctt
      421 gtactcgtcg gtgatcacgg cccagcccac agagttggtg ccgatgtcca ggccgatgct
      481 gtacttcttg tcggctgctg ggactccgtg gataccgacc ttccgcttct tctttggggc
      541 catcttatcg tcatcgtctt tgtaatcaat atcatgatcc ttgtagtctc cgtcgtggtc
      601 cttatagtcc atggtggcac cggtccaacc tgaaaaaaag tgatttcagg caggtgctcc
      661 aggtaattaa acattaatac cccaccaacc aaccatccct taaaccctta cctcttgctc
      721 agctaattac agcccggagg agaagggccg tcccgcccgc tcacctgtgg gagtaacgcg
      781 gtcagtcaga gccggggcgg gcggcgcgag gcggcggcgg agcggggcac ggggcgaagg
      841 cagcgcgcag cgactcccgc ccgccgcgcg cttcgctttt tatagggccg ccgccgccgc
      901 cgcctcgcca taaaaggaaa ctttcggagc gcgccgctct gattggctgc cgccgcacct
      961 ctccgcctcg ccccgccccg cccctcgccc cgccccgccc cgcctggcgc gcgccccccc
     1021 cccccccccc gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg
     1081 gggtggggag gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat
     1141 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     1201 gtactgggca caatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     1261 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     1321 ttgacgtcaa tggaaagtcc ctattggcgt tactattgac gtcaatgggc gggggtcgtt
     1381 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgggtacct ctagagccat
     1441 ttgtctgcag aattggcgca cgcgctaaaa acggactagc cttattttaa cttgctattt
     1501 ctagctctaa aacaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta
     1561 gccttatttt aacttgctat ttctagctct aaaacctagg gacaggattg gtgaccggtg
     1621 tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca agttacggta
     1681 agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat
     1741 tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcca
     1801 attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg
     1861 ccctcacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2761 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2821 accgcggctt ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2881 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2941 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3001 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3061 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3121 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3181 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3241 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3301 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3361 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3421 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3481 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3541 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3601 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3661 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3721 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3781 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3841 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3901 atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa
     3961 attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa
     4021 aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa
     4081 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4141 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     4201 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4261 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc
     4321 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4381 gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg
     4441 agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     4501 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     4561 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg ctccccagca
     4621 tgcctgctat tctcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     4681 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     4741 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     4801 tggcaactag aaggcacagt cgaggctgat cagcgagctc taggaattcc tacttgtaca
     4861 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg
     4921 tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca
     4981 gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt
     5041 ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     5101 gctcggagga ggcctcccag cccatggttt tcttctgcat tacggggccg tcggagggga
     5161 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     5221 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     5281 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     5341 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     5401 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     5461 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     5521 tgatgatggc catgttatcc tcctcgccct tgctcaccat ggtggcgacc ggcatgaatt
     5581 caggtccagg gttctcctcc acgtctccag cctgcttcag caggctgaag ttagtagctc
     5641 cgcttcccag cgcctttctc cgtttttctg ccacttcctt ccaatactga gaggatggat
     5701 tttctttaat cataagatca aatgactcct gggtgactcc tccaagattt ttattttcac
     5761 tagattctgg gacaataacc ccagggctgg aagttgtaga tgttaagtgg tcattccgat
     5821 gtttcctttt ggacaagcct gcggacagct cattttctct tccaacaaga gatccagatg
     5881 cagaaggctg aatcatcttc agagttcttc ttgggacaga actattcttt atattctctt
     5941 tgatttcttc ttgtttctgc ttcatactgg gattcataag cttggatccc tttttctttt
     6001 ttgcctggcc ggcctttttc gtggccgccg gccttttgtc gcctcccagc tgagacaggt
     6061 cgatccgtgt ctcgtacagg ccggtgatgc tctggtggat cagggtggcg tccagcacct
     6121 ctttggtgct ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg aaggcggcag
     6181 gggctcccag attggtcagg gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg
     6241 gcttatcccg gtgcttgttg taggcggaca gcactttgtc cagattagcg tcggccagga
     6301 tcactctctt ggagaactcg ctgatctgct cgatgatctc gtccaggtag tgcttgtgct
     6361 gttccacaaa cagctgtttc tgctcattat cctcggggga gcccttcagc ttctcatagt
     6421 ggctggccag gtacaggaag ttcacatatt tggagggcag ggccagttcg tttcccttct
     6481 gcagttcgcc ggcagaggcc agcattctct tccggccgtt ttccagctcg aacagggagt
     6541 acttaggcag cttgatgatc aggtcctttt tcacttcttt gtagcccttg gcttccagaa
     6601 agtcgatggg attcttctcg aagctgcttc tttccatgat ggtgatcccc agcagctctt
     6661 tcacactctt cagtttcttg gacttgccct tttccacttt ggccaccacc agcacagaat
     6721 aggccacggt ggggctgtcg aagccgccgt acttcttagg gtcccagtcc ttctttctgg
     6781 cgatcagctt atcgctgttc ctcttgggca ggatagactc tttgctgaag ccgcctgtct
     6841 gcacctcggt ctttttcacg atattcactt ggggcatgct cagcactttc cgcacggtgg
     6901 caaaatcccg gcccttatcc cacacgatct ccccggtttc gccgtttgtc tcgatcagag
     6961 gccgcttccg gatctcgccg ttggccaggg taatctcggt cttgaaaaag ttcatgatgt
     7021 tgctgtagaa gaagtacttg gcggtagcct tgccgatttc ctgctcgctc ttggcgatca
     7081 tcttccgcac gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt
     7141 actttttgat cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt
     7201 agttgttgat ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct
     7261 tggacttcag ggtgatcact ttcacttccc ggatcagctt gtcattctcg tcgtacttag
     7321 tgttcatccg ggagtccagg atctgtgcca cgtgctttgt gatctgccgg gtttccacca
     7381 gctgtctctt gatgaagccg gccttatcca gttcgctcag gccgcctctc tcggccttgg
     7441 tcagattgtc gaactttctc tgggtaatca gcttggcgtt cagcagctgc cgccagtagt
     7501 tcttcatctt cttcacgacc tcttcggagg gcacgttgtc gctcttgccc cggttcttgt
     7561 cgcttctggt cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga
     7621 tatggtccac atcgtagtcg gacagccggt tgatgtccag ttcctggtcc acgtacatat
     7681 cccgcccatt ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca
     7741 cggggtgttc tttcaggatc tggctgccca gctctttgat gccctcttcg atccgcttca
     7801 ttctctcgcg gctgttcttc tgtcccttct gggtggtctg gttctctctg gccatttcga
     7861 tcacgatgtt ctcgggcttg tgccggccca tcactttcac gagctcgtcc accaccttca
     7921 ctgtctgcag gatgcccttc ttaatggcgg ggctgccggc cagattggca atgtgctcgt
     7981 gcaggctatc gccctggccg gacacctggg ctttctggat gtcctcttta aaggtcaggc
     8041 tgtcgtcgtg gatcagctgc atgaagtttc tgttggcgaa gccgtcggac ttcaggaaat
     8101 ccaggattgt cttgccggac tgcttgtccc ggatgccgtt gatcagcttc cggctcagcc
     8161 tgccccagcc ggtgtatctc cgccgcttca gctgcttcat cactttgtcg tcgaacaggt
     8221 gggcataggt tttcagccgt tcctcgatca tctctctgtc ctcaaacagt gtcagggtca
     8281 gcacgatatc ttccagaatg tcctcgtttt cctcattgtc caggaagtcc ttgtccttga
     8341 taattttcag cagatcgtgg tatgtgccca gggaggcgtt gaaccgatct tccacgccgg
     8401 agatttccac ggagtcgaag cactcgattt tcttgaagta gtcctctttc agctgcttca
     8461 cggtcacttt ccggttggtc ttgaacagca ggtccacgat ggcctttttc tgctcgccgc
     8521 tcaggaaggc gggctttctc attccctcgg tcacgtattt cactttggtc agctcgttat
     8581 acacggtgaa gtactcgtac agcaggctgt gcttgggcag caccttctcg ttgggcaggt
     8641 tcttatcgaa gttggtcatc cgctcgatga agctctgggc ggaagcgccc ttgtccacca
     8701 cttcctcgaa gttccagggg gtgatggttt cctcgctctt tctggtcatc caggcgaatc
     8761 tgctgtttcc cctggccaga gggcccacgt agtaggggat gcggaaggtc aggatcttct
     8821 cgatcttttc ccggttgtcc ttcaggaatg ggtaaaaatc ttcctgccgc cgcagaatgg
     8881 cgtgcagctc tcccaggtgg atctggtggg ggatgctgcc gttgtcgaag gtccgctgct
     8941 tccgcagcag gtcctctctg ttcagcttca cgagcagttc ctcggtgccg tccatctttt
     9001 ccaggatggg cttgatgaac ttgtagaact cttcctggct ggctccgccg tcaatgtagc
     9061 cggcgtagcc gttcttgctc tggtcgaaga aaatctcttt gtacttctca ggcagctgct
     9121 gccgcacgag agctttcagc agggtcaggt cctggtggtg ctcgtcgtat ctcttgatca
     9181 tagaggcgct caggggggcc ttggtgatct cggtgttcac tctcaggatg tcgctcagca
     9241 ggatggcgtc ggacaggttc ttggcggcca gaaacaggtc ggcgtactgg tcgccgatct
     9301 gggccagcag gttgtccagg tcgtcgtcgt aggtgtcctt gctcagctgc agtttggcat
     9361 cctcggccag gtcgaagttg ctcttgaagt tgggggtcag gcccaggctc agggcaatca
     9421 ggtttccgaa caggccattc ttcttctcgc cgggcagctg ggcgatcaga ttttccagcc
     9481 gtctgctctt gctcagtctg gcagacagga tggccttggc gtccacgccg ctggcgttga
     9541 tggggttttc ctcgaacagc tggttgtagg tctgcaccag ctggatgaac agcttgtcca
     9601 cgtcgctgtt gtcggggttc aggtcgccct cgatcaggaa gtggcc
//
