LOCUS       sequence-407168-       10292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3XHA tagged dead Cas9 from F. novicida with KRAB repressor domain, 
            T2A self-cleaving peptide and enhanced GFP biomarker for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10292)
  TITLE     3XHA tagged dead Cas9 from F. novicida with KRAB repressor domain, 
            T2A self-cleaving peptide region and enhanced GFP biomarker for 
            mammalian expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10292)
  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            "Unpublished"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: PX458_3XHA_dFnCas9_KRAB_T2A_eGFP
FEATURES             Location/Qualifiers
     source          1..10292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..276
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     28..48
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     199..218
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        295..367
                     /label=Fn gRNA scaffold
                     /note="guide RNA scaffold for the Francisella novicida 
                     CRISPR/Cas9 system (Hirano et al., 2016)"
     enhancer        464..749
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1001..1228
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1240..1249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1255..1335
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             1342..1362
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1387..6270
                     /codon_start=1
                     /product="Cas9 endonuclease from the Francisella novicida 
                     Type II CRISPR/Cas system (Hirano et al., 2016)"
                     /label=FnCas9
                     /translation="NFKILPIAIALGVKNTGVFSAFYQKGTSLERLDNKNGKVYELSKD
                     SYTLLMNNRTARRHQRRGIDRKQLVKRLFKLIWTEQLNLEWDKDTQQAISFLFNRRGFS
                     FITDGYSPEYLNIVPEQVKAILMDIFDDYNGEDDLDSYLKLATEQESKISEIYNKLMQK
                     ILEFKLMKLCTDIKDDKVSTKTLKEITSYEFELLADYLANYSESLKTQKFSYTDKQGNL
                     KELSYYHHDKYNIQEFLKRHATINDRILDTLLTDDLDIWNFNFEKFDFDKNEEKLQNQE
                     DKDHIQAHLHHFVFAVNKIKSEMASGGRHRSQYFQEITNVLDENNHQEGYLKNFCENLH
                     NKKYSNLSVKNLVNLIGNLSNLELKPLRKYFNDKIHAKADHWDEQKFTETYCHWILGEW
                     RVGVKDQDKKDGAKYSYKDLCNELKQKVTKAGLVDFLLELDPCRTIPPYLDNNNRKPPK
                     CQSLILNPKFLDNQYPNWQQYLQELKKLQSIQNYLDSFETDLKVLKSSKDQPYFVEYKS
                     SNQQIASGQRDYKDLDARILQFIFDRVKASDELLLNEIYFQAKKLKQKASSELEKLESS
                     KKLDEVIANSQLSQILKSQHTNGIFEQGTFLHLVCKYYKQRQRARDSRLYIMPEYRYDK
                     KLHKYNNTGRFDDDNQLLTYCNHKPRQKRYQLLNDLAGVLQVSPNFLKDKIGSDDDLFI
                     SKWLVEHIRGFKKACEDSLKIQKDNRGLLNHKINIARNTKGKCEKEIFNLICKIEGSED
                     KKGNYKHGLAYELGVLLFGEPNEASKPEFDRKIKKFNSIYSFAQIQQIAFAERKGNANT
                     CAVCSADNAHRMQQIKITEPVEDNKDKIILSAKAQRLPAIPTRIVDGAVKKMATILAKN
                     IVDDNWQNIKQVLSAKHQLHIPIITESNAFEFEPALADVKGKSLKDRRKKALERISPEN
                     IFKDKNNRIKEFAKGISAYSGANLTDGDFDGAKEELDAIIPRSHKKYGTLNDEANLICV
                     TRGDNKNKGNRIFCLRDLADNYKLKQFETTDDLEIEKKIADTIWDANKKDFKFGNYRSF
                     INLTPQEQKAFRHALFLADENPIKQAVIRAINNRNRTFVNGTQRYFAEVLANNIYLRAK
                     KENLNTDKISFDYFGIPTIGNGRGIAEIRQLYEKVDSDIQAYAKGDKPQASYSHLIDAM
                     LAFCIAADEHRNDGSIGLEIDKNYSLYPLDKNTGEVFTKDIFSQIKITDNEFSDKKLVR
                     KKAIEGFNTHRQMTRDGIYAENYLPILIHKELNEVRKGYTWKNSEEIKIFKGKKYDIQQ
                     LNNLVYCLKFVDKPISIDIQISTLEELRNILTTNNIAATAEYYYINLKTQKLHEYYIEN
                     YNTALGYKKYSKEMEFLRSLAYRSERVKIKSIDDVKQVLDKDSNFIIGKITLPFKKEWQ
                     RLYREWQNTTIKDDYEFLKSFFNVKSITKLHKKVRKDFSLPISTNEGKFLVKRKTWDNN
                     FIYQILNDSDSRADGTKPFIPAFDISKNEIVEAIIDSFTSKNIFWLPKNIELQKVDNKN
                     IFAIDTSKWFEVETPSDLRDIGIATIQYKIDNNSRPKVRVKLDYVIDDDSKINYFMNHS
                     LLKSRYPDKVLEILKQSTIIEFESSGFNKTIKEMLGMKLAGIYNETSNN"
     CDS             6271..6318
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6364..6549
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             6565..6618
                     /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(6619..6621,6622..7332)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6619..6621,6622..7332)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6661..6682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6922..6941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7269..7290
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7360..7377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7366..7573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7582..7722
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7582..7711
                     /label=AAV2 ITR
     rep_origin      7797..8252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7884..7903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8094..8115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8269..8288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8388..8410
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8448..8466)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8534..8638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8639..8707,8708..9499)
                     /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(8857..8876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9670..10258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10159..10178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctg gccttttgct cacatgtgag ggcctatttc ccatgattcc ttcatatttg
       61 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
      121 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      181 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
      241 ttggctttat atatcttgtg gaaaggacga aacaccgggt cttcgagaag acctgtttca
      301 gttgcgccga aaggcgctct gtaatcattt aaaagtattt tgaacggacc tctgtttgac
      361 acgtctgttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt
      421 agcgcgtgcg ccaattctgc agacaaatgg ctctagaggt acccgttaca taacttacgg
      481 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc
      541 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      601 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      661 ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      721 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
      781 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
      841 gtgcagcgat gggggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
      901 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      961 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     1021 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     1081 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag
     1141 caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga
     1201 gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatggg atcctaccca
     1261 tacgatgttc cagattacgc ttacccatac gatgttccag attacgctta cccatacgat
     1321 gttccagatt acgctatggc cccaaagaag aagcggaagg tcagtatcca cggagtccca
     1381 gcagccaatt ttaagatcct gcctatcgca atcgccctgg gcgtcaagaa tactggcgtg
     1441 tttagtgctt tttatcagaa ggggacctca ctggagagac tggacaataa gaacggaaaa
     1501 gtgtatgaac tgtccaagga ttcttacact ctgctgatga acaataggac cgcacggaga
     1561 caccagaggc gaggaattga caggaaacag ctggtgaagc gcctgttcaa actgatctgg
     1621 acagagcagc tgaacctgga atgggataag gacactcagc aggccatcag cttcctgttt
     1681 aatcgacggg gattctcttt tattactgac ggctatagtc ctgagtacct gaacatcgtg
     1741 ccagaacagg tcaaggcaat cctgatggac attttcgacg attataatgg cgaggacgat
     1801 ctggattcct acctgaaact ggccacagag caagagagta agatcagcga aatctacaac
     1861 aagctgatgc agaagatcct ggagttcaag ctgatgaaac tgtgcaccga catcaaggac
     1921 gataaagtga gtaccaagac actgaaagag atcacaagct acgagttcga actgctggcc
     1981 gattatctgg ctaactacag cgaatccctg aaaacccaga aattttccta cacagacaag
     2041 cagggcaatc tgaaagagct gtcttactac caccatgata agtacaacat ccaggagttc
     2101 ctgaagagac acgccaccat caatgacagg attctggata cactgctgac tgacgatctg
     2161 gacatctgga acttcaactt cgagaagttc gatttcgaca agaacgagga aaaactgcag
     2221 aatcaggaag ataaggacca cattcaggct catctgcacc atttcgtgtt tgcagtcaat
     2281 aagatcaaaa gcgagatggc atccggcggg cgccatcgaa gccagtactt ccaggaaatc
     2341 accaacgtgc tggacgagaa caatcaccag gaaggctacc tgaaaaactt ctgtgagaat
     2401 ctgcataaca agaagtacag caatctgtcc gtgaagaatc tggtcaacct gattggaaat
     2461 ctgtccaacc tggaactgaa gcccctgcgc aaatacttca acgacaagat ccacgctaaa
     2521 gcagaccatt gggatgagca gaagtttact gaaacctatt gccactggat tctgggcgag
     2581 tggcgggtgg gggtcaagga tcaggacaag aaagacggcg caaagtattc ttacaaggac
     2641 ctgtgtaacg agctgaagca gaaagtgact aaggccgggc tggtggactt cctgctggag
     2701 ctggacccct gccgaaccat tccaccttac ctggacaaca ataacagaaa gccacccaaa
     2761 tgtcagagcc tgatcctgaa tcccaagttt ctggataatc agtatcctaa ctggcagcag
     2821 tacctgcagg agctgaagaa actgcagtca atccagaact acctggacag cttcgaaacc
     2881 gatctgaagg tgctgaaaag ctccaaggac cagccttact tcgtcgagta caagtctagt
     2941 aaccagcaga tcgcttccgg ccagcgggat tacaaggatc tggacgcaag aatcctgcag
     3001 ttcatttttg acagggtgaa ggcctctgat gagctgctgc tgaacgaaat ctatttccag
     3061 gcaaagaaac tgaagcagaa agcctcaagc gagctggaaa agctggagtc ctctaagaaa
     3121 ctggacgaag tgatcgctaa ctctcagctg agtcagattc tgaagtctca gcacacaaat
     3181 ggaatcttcg agcagggcac ttttctgcat ctggtgtgca aatactataa gcagcgacag
     3241 agagccaggg acagccgcct gtacatcatg cctgaatatc gatacgataa gaaactgcac
     3301 aagtacaaca acaccggccg ctttgacgat gacaaccagc tgctgacata ttgtaatcat
     3361 aagccccggc agaaaagata ccagctgctg aacgacctgg caggagtgct gcaggtctct
     3421 cctaattttc tgaaggataa aatcgggtcc gatgacgatc tgttcatttc taagtggctg
     3481 gtggagcaca tccggggctt taagaaggcc tgcgaagata gcctgaaaat ccagaaggat
     3541 aacaggggac tgctgaatca taagatcaac attgcacgca ataccaaggg caaatgcgag
     3601 aaagaaatct tcaacctgat ctgtaagatt gaggggagcg aagataagaa agggaattat
     3661 aagcacggac tggcctacga gctgggagtg ctgctgttcg gagagccaaa cgaggccagc
     3721 aagcccgaat ttgataggaa aatcaagaaa ttcaattcaa tctacagctt tgcccagatc
     3781 cagcagattg cctttgctga gaggaagggg aatgcaaaca catgcgccgt gtgtagtgca
     3841 gacaacgccc atcgcatgca gcagatcaaa attactgagc cagtcgaaga taataaggat
     3901 aaaatcattc tgtcagcaaa ggcacagcga ctgcctgcaa tcccaacccg aattgtggat
     3961 ggagctgtca agaaaatggc tacaattctg gcaaagaata tcgtggacga taattggcag
     4021 aacattaagc aggtcctgag cgcaaaacac cagctgcata tcccaatcat taccgagtcc
     4081 aacgccttcg agtttgaacc cgctctggca gacgtgaagg gcaaatctct gaaggataga
     4141 aggaagaaag ccctggagcg aattagtccc gaaaacatct tcaaggataa gaacaacaga
     4201 atcaaggagt ttgctaaggg gatttccgcc tactctggag ctaacctgac agatggggac
     4261 ttcgatggag caaaggagga actggatgcc atcattcctc gcagccataa gaaatatggc
     4321 actctgaacg acgaggctaa tctgatttgc gtgacccggg gcgataataa gaacaaaggg
     4381 aaccggatct tctgtctgag agacctggcc gataattaca agctgaaaca gtttgagacc
     4441 acagacgatc tggagatcga aaagaaaatt gccgacacca tctgggatgc taataagaag
     4501 gacttcaagt tcggaaacta tcggagcttc atcaatctga cacctcagga gcagaaagca
     4561 ttcagacacg ccctgtttct ggctgatgaa aacccaatca agcaggcagt gatcagagcc
     4621 attaataacc gcaaccgaac cttcgtgaat ggcacacaga ggtattttgc tgaggtcctg
     4681 gcaaataaca tctacctgcg cgccaagaaa gaaaatctga acactgacaa gatcagcttc
     4741 gattactttg gaatccctac cattggaaac ggccgaggga tcgctgagat tcggcagctg
     4801 tatgaaaagg tggacagtga tatccaggcc tacgctaaag gcgacaagcc acaggcctct
     4861 tatagtcacc tgattgatgc tatgctggca ttctgcatcg ccgctgacga gcatcggaac
     4921 gatggatcta ttggcctgga aatcgacaaa aactatagtc tgtaccctct ggataagaat
     4981 actggcgagg tgttcaccaa agacatcttt tcacagatca agattaccga caacgagttc
     5041 agcgataaga aactggtcag aaagaaagct attgaagggt ttaacacaca cagacagatg
     5101 actagggatg gaatctatgc agagaattac ctgcctatcc tgattcataa ggagctgaac
     5161 gaagtgagga aggggtacac atggaaaaat tccgaggaaa tcaaaatttt caagggaaag
     5221 aaatacgaca tccagcagct gaataacctg gtgtattgtc tgaagtttgt ggacaaacca
     5281 atcagtattg atatccagat ttcaaccctg gaggaactga gaaacatcct gactaccaat
     5341 aacattgcag ccactgccga gtactattac attaatctga aaacccagaa gctgcacgag
     5401 tattacatcg aaaattacaa cacagccctg gggtataaga aatacagcaa ggagatggag
     5461 ttcctgaggt ccctggctta taggtctgag cgcgtgaaga tcaaaagtat tgacgatgtc
     5521 aagcaggtcc tggacaagga ttcaaacttc atcatcggaa agatcacact gcccttcaag
     5581 aaagagtggc agcgactgta ccgggaatgg cagaacacaa ctatcaaaga cgattatgag
     5641 tttctgaaga gcttctttaa tgtgaagtcc attactaaac tgcacaagaa agtccggaaa
     5701 gacttctctc tgcccatcag tacaaacgag ggcaagtttc tggtgaagag aaaaacttgg
     5761 gataataact tcatctacca gattctgaat gactcagata gcagggcaga cgggactaaa
     5821 cccttcattc ctgcctttga tatcagcaag aacgagattg tggaagccat cattgacagt
     5881 ttcacctcaa aaaacatctt ttggctgcca aagaatattg agctgcagaa ggtggacaac
     5941 aagaacatct tcgccattga taccagcaag tggtttgagg tcgaaacacc atccgacctg
     6001 cgcgatatcg gcattgctac cattcagtac aagatcgaca ataactcacg ccccaaggtg
     6061 cgagtcaaac tggattacgt gatcgacgat gacagcaaga ttaactattt catgaatcac
     6121 tcactgctga agagccggta tcccgacaaa gtcctggaga tcctgaagca gagcacaatc
     6181 attgagttcg aaagttcagg gtttaacaaa actattaagg agatgctggg aatgaagctg
     6241 gccggcatct acaatgaaac ctccaataac aaaaggccgg cggccacgaa aaaggccggc
     6301 caggcaaaaa agaaaaaggg cggtggcgga gggatggatg ctaagtcact aactgcctgg
     6361 tcccggacac tggtgacctt caaggatgta tttgtggact tcaccaggga ggagtggaag
     6421 ctgctggaca ctgctcagca gatcgtgtac agaaatgtga tgctggagaa ctataagaac
     6481 ctggtttcct tgggttatca gcttactaag ccagatgtga tcctccggtt ggagaaggga
     6541 gaagagcccg aattcggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc
     6601 gaggagaatc ctggcccagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     6661 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     6721 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     6781 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     6841 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     6901 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     6961 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7021 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     7081 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     7141 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     7201 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     7261 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     7321 gagctgtaca aggagttcta actagagctc gctgatcagc ctcgactgtg ccttctagtt
     7381 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     7441 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7501 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     7561 ggcatgctgg ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     7621 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     7681 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     7741 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     7801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     7861 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     7921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     7981 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     8041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     8101 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     8161 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     8221 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     8281 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     8341 ggcttgtctg ctcccagcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     8401 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     8461 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     8521 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     8581 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     8641 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     8701 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     8761 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     8821 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     8881 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     8941 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     9001 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     9061 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     9121 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     9181 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     9241 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     9301 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
     9361 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     9421 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     9481 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     9541 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     9601 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     9661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     9721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     9781 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     9841 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     9901 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     9961 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    10021 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    10081 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    10141 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    10201 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    10261 cgccagcaac gcggcctttt tacggttcct gg
//
