LOCUS       sequence-224718-        9604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-GFP, dual gRNA expression vector for targeted integration of 
            repair templates into safe harbor 231 locus in chromosome 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9604)
  AUTHORS   Pellenz S, Phelps M, Tang W, Hovde BT, Sinit RB, Fu W, Li H, Chen E,
            Monnat RJ Jr
  TITLE     New Human Chromosomal Sites with ""Safe Harbor"" Potential for 
            Targeted Transgene Insertion.
  JOURNAL   Hum Gene Ther. 2019 Jul;30(7):814-828. doi: 10.1089/hum.2018.169. 
            Epub 2019 Mar 28.
  PUBMED    30793977
REFERENCE   2  (bases 1 to 9604)
  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"; doi: 
            "10.1089/hum.2018.169"; journalName: "Hum Gene Ther"; date: 
            "2019-07"; volume: "30"; issue: "7"; pages: "814-828"
COMMENT     Sequence Label: pUS2-SH231
FEATURES             Location/Qualifiers
     source          1..9604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..298
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        318..393
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        426..674
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        695..770
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        799..1084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     regulatory      1603..1612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1612..1677
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1684..1704
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1729..5829
                     /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             1729..5829
                     /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             5830..5877
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5884..5937
                     /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(5944..5946,5947..5949,5950..6663)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6250..6269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6597..6618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6691..6708)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6697..6904
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6913..7053
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6913..7042
                     /label=AAV2 ITR
     rep_origin      7128..7583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7215..7234)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7425..7446
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7600..7619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7719..7741
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7779..7797)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7865..7969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7970..8038,8039..8830)
                     /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(8188..8207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9001..9589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9490..9509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttgtg agggcctatt
       61 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      121 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      181 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      241 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgc
      301 ctcccccata gtaccatgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      361 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg accaacgagc aagataagga
      421 ctagggaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
      481 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      541 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      601 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      661 aaggacgaaa caccgatgtg ctcactgagt ctgagtttta gagctagaaa tagcaagtta
      721 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttctcg
      781 agaaactagt aaggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      841 caacgacccc cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca
      901 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      961 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta
     1021 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1081 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
     1141 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
     1201 gggggggcgc gcgccrggsg gggggggggg gggggggggg gggggggggg gggggsggrg
     1261 rggkgsggcg gcrgccaatc agrgcggcgc gctccgaaag tttcctttta tggcgrggcg
     1321 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggrgtcg ctgcgcgctg
     1381 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
     1441 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct
     1501 gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct
     1561 ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag
     1621 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
     1681 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
     1741 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     1801 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     1861 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     1921 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     1981 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     2041 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     2101 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     2161 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     2221 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     2281 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     2341 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     2401 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     2461 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     2521 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     2581 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     2641 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     2701 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     2761 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     2821 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     2881 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     2941 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     3001 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     3061 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     3121 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     3181 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     3241 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     3301 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     3361 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     3421 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     3481 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     3541 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     3601 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     3661 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     3721 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     3781 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     3841 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     3901 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     3961 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     4021 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     4081 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     4141 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     4201 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     4261 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     4321 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     4381 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     4441 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     4501 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     4561 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     4621 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     4681 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     4741 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     4801 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     4861 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     4921 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     4981 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     5041 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     5101 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     5161 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     5221 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     5281 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     5341 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     5401 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     5461 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     5521 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     5581 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     5641 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     5701 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     5761 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     5821 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggct
     5881 agcgagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaacc
     5941 ggtatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     6001 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     6061 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     6121 accctcgtga ccaccttcac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     6181 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     6241 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     6301 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     6361 cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag
     6421 aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc
     6481 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     6541 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     6601 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     6661 taagaattct aactagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     6721 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6781 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6841 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     6901 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     6961 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     7021 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     7081 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     7141 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7201 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7261 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7321 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     7381 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7441 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     7501 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7561 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     7621 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     7681 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     7741 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     7801 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7921 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7981 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     8041 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     8101 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     8161 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     8221 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     8281 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     8341 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     8401 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8461 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8521 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8581 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8641 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     8701 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8761 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8821 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     9001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     9061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     9121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     9181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     9241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     9301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     9361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     9421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     9481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     9541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     9601 cgcg
//
