LOCUS       sequence-211963-        9825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA cassette (BsmBI cloning site) and EF1-A driven 
            eSpCas9 linked via P2A with mRuby2 and dominant negative mtp53 for 
            enhanced survival of hES after DSB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9825)
  AUTHORS   Ang LT, Tan AKY, Autio MI, Goh SH, Choo SH, Lee KL, Tan J, Pan B, 
            Lee JJH, Lum JJ, Lim CYY, Yeo IKX, Wong CJY, Liu M, Oh JLL, Chia 
            CPL, Loh CH, Chen A, Chen Q, Weissman IL, Loh KM, Lim B
  TITLE     A Roadmap for Human Liver Differentiation from Pluripotent Stem 
            Cells.
  JOURNAL   Cell Rep. 2018 Feb 20;22(8):2190-2205. doi: 
            10.1016/j.celrep.2018.01.087.
  PUBMED    29466743
REFERENCE   2  (bases 1 to 9825)
  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: 
            "10.1016/j.celrep.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-20- 20"; volume: "22"; issue: "8"; pages: "2190-2205"
COMMENT     Sequence Label: pMIA3
FEATURES             Location/Qualifiers
     source          1..9825
                     /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)"
     primer_bind     50..70
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     221..240
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        315..390
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        503..714
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          715..1653
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1610..1630
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1694..1703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1703..1768
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1775..1795
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1820..4357,4358..4360,4361..4822,4823..4825,4826..4993,
                     4994..4996,4997..5920)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5924..5944
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5963..6019
                     /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             6020..6730
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     CDS             6740..6796
                     /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"
     primer_bind     complement(7133..7150)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7139..7346
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7355..7495
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7355..7484
                     /label=AAV2 ITR
     rep_origin      7570..7998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7657..7676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7867..7888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8042..8146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8147..8215,8216..9007)
                     /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(8365..8384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9178..9766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9667..9686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgggccca gccggccaga tctgagctcg cggccgcgat atcgctagcg 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 gaaacaccgg
      301 agacggacgt ctctgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
      361 caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt
      421 taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga
      481 ttcactcgag gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
      541 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
      601 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
      661 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
      721 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
      781 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      841 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      901 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      961 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
     1021 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
     1081 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
     1141 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
     1201 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
     1261 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     1321 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     1381 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     1441 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1501 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1561 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1621 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggataatc actttttttc
     1681 aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga
     1741 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
     1801 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     1861 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     1921 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     1981 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     2041 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     2101 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     2161 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     2221 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     2281 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     2341 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     2401 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     2461 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     2521 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     2581 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     2641 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     2701 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     2761 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     2821 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     2881 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     2941 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     3001 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     3061 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     3121 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     3181 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     3241 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     3301 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     3361 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     3421 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     3481 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     3541 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     3601 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     3661 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     3721 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     3781 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     3841 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     3901 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     3961 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     4021 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     4081 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     4141 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     4201 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     4261 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     4321 ctacgatgtg gaccatatcg tgcctcagag ctttctggcc gacgactcca tcgacaacaa
     4381 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     4441 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     4501 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     4561 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     4621 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     4681 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     4741 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     4801 cgccctgatc aaaaagtacc ctgccctgga aagcgagttc gtgtacggcg actacaaggt
     4861 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     4921 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     4981 cgagatccgg aaggcccctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     5041 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     5101 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     5161 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     5221 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     5281 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     5341 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     5401 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     5461 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     5521 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     5581 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     5641 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     5701 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     5761 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     5821 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     5881 gtacgagaca cggatcgacc tgtctcagct gggaggcgac agccccaaga agaagagaaa
     5941 ggtggaggcc agcggaagcg gagccactaa cttctccctg ttgaaacaag caggggatgt
     6001 cgaagagaat cccgggccaa tggtgtctaa gggcgaggag ctgatcaagg aaaatatgcg
     6061 tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg
     6121 agaaggcaat ccgtacatgg gaactcaaac catgaggatc aaagtcatcg agggaggacc
     6181 cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc gtacttttat
     6241 caagtacccg aaaggcattc ctgatttctt taaacagtcc tttcctgagg gttttacttg
     6301 ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcatgcagg acaccagcct
     6361 tgaggatggc tgtctcgttt accacgtcca agtcagaggg gtaaactttc cctccaatgg
     6421 tcccgtgatg cagaaaaaga ccaagggttg ggagcctaat acagagatga tgtatccagc
     6481 agatggtggt ctgaggggat acactcatat ggcactgaaa gttgatggtg gtggccatct
     6541 gtcttgctct ttcgtaacaa cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc
     6601 cggtatccat gccgttgatc accgcctgga aaggttagag gaaagtgaca atgaaatgtt
     6661 cgtagtacaa cgcgaacacg cagttgccaa gttcgccggg cttggtggtg ggatggacga
     6721 gctgtacaag ggaagcggag ccactaactt ctccctgttg aaacaagcag gggatgtcga
     6781 agagaatccc gggccaatga ctgccatgga ggagtcacag tcggatatca gcctcaagag
     6841 agcgctgccc acctgcacaa gcgcctctcc cccgcaaaag aaaaaaccac ttgatggaga
     6901 gtatttcacc ctcaagatcc gcgggcgtaa acgcttcgag atgttccggg agctgaatga
     6961 ggccttagag ttaaaggatg cccatgctac agaggagtct ggagacagca gggctcactc
     7021 cagctacctg aagaccaaga agggccagtc tacttcccgc cataaaaaaa caatggtcaa
     7081 gaaagtgggg cctgactcag actgataaga attcctagag ctcgctgatc agcctcgact
     7141 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     7201 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     7261 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     7321 gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc
     7381 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     7441 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     7501 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     7561 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     7621 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     7681 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     7741 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     7801 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     7861 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     7921 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     7981 aaaaatttaa cgcgaatttt aacaaaatta gacgtcaggt ggcacttttc ggggaaatgt
     8041 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     8101 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     8161 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     8221 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     8281 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     8341 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     8401 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     8461 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     8521 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     8581 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     8641 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     8701 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     8761 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8821 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8881 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8941 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     9001 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     9061 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     9121 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     9181 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     9241 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     9301 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     9361 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     9421 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     9481 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     9541 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     9601 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     9661 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     9721 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9781 ggccttttta cggttcctgg ccttttgctg gccttttgct cacat
//
