LOCUS       sequence-433843-        9829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eSpCas9 and gRNA targeting Pansio-1 GSH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9829)
  AUTHORS   Autio MI, Motakis E, Perrin A, Bin Amin T, Tiang Z, Do DV, Wang J, 
            Tan JKM, Ding SSL, Tan WX, Lee CJM, Teo AKK, Foo R
  TITLE     Computationally defined and in vitro validated putative genomic safe
            harbour loci for transgene expression in human cells.
  JOURNAL   Elife. 2024 Jan 2;13:e79592. doi: 10.7554/eLife.79592.
  PUBMED    38164941
REFERENCE   2  (bases 1 to 9829)
  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: "Elife."; 
            date: "2024-01-2"; volume: "13:e79592. doi"; pages: " 
            10.7554/eLife.79592"
COMMENT     Sequence Label: pMIA3-Pansio
FEATURES             Location/Qualifiers
     source          1..9829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..287
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     39..59
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     210..229
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        308..383
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        496..707
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          708..1646
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1603..1623
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1687..1696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1696..1761
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1768..1788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1813..4350,4351..4353,4354..4815,4816..4818,4819..4986,
                     4987..4989,4990..5913)
                     /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             5917..5937
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5956..6012
                     /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             6013..6723
                     /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             6733..6789
                     /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(7126..7143)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7132..7339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7348..7488
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7348..7477
                     /label=AAV2 ITR
     rep_origin      7563..7991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7650..7669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7860..7881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8035..8139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8140..8208,8209..9000)
                     /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(8358..8377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9171..9759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9660..9679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccggccagat ctgagctcgc ggccgcgata tcgctagcga gggcctattt cccatgattc
       61 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
      121 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
      181 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
      241 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgca cctggagaca
      301 ctaccaagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      361 aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata
      421 aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agattcactc
      481 gaggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      541 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      601 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      661 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgctgtgt
      721 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      781 ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
      841 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
      901 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
      961 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
     1021 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
     1081 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
     1141 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
     1201 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     1261 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
     1321 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     1381 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1441 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     1501 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     1561 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     1621 ttcaaagttt ttttcttcca tttcaggtgt cgtgaggata atcacttttt ttcaggttgg
     1681 accggtgcca ccatggacta taaggaccac gacggagact acaaggatca tgatattgat
     1741 tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga
     1801 gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     1861 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     1921 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     1981 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     2041 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     2101 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     2161 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     2221 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     2281 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     2341 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     2401 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     2461 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     2521 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     2581 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     2641 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     2701 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     2761 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     2821 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     2881 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     2941 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     3001 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     3061 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     3121 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     3181 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     3241 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     3301 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     3361 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     3421 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     3481 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     3541 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     3601 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     3661 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     3721 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     3781 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     3841 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     3901 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     3961 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     4021 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     4081 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     4141 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     4201 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     4261 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     4321 gtggaccata tcgtgcctca gagctttctg gccgacgact ccatcgacaa caaggtgctg
     4381 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     4441 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     4501 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     4561 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     4621 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     4681 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     4741 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     4801 atcaaaaagt accctgccct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     4861 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     4921 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     4981 cggaaggccc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     5041 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     5101 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     5161 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     5221 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     5281 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     5341 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     5401 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     5461 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     5521 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     5581 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     5641 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     5701 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     5761 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     5821 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     5881 acacggatcg acctgtctca gctgggaggc gacagcccca agaagaagag aaaggtggag
     5941 gccagcggaa gcggagccac taacttctcc ctgttgaaac aagcagggga tgtcgaagag
     6001 aatcccgggc caatggtgtc taagggcgag gagctgatca aggaaaatat gcgtatgaag
     6061 gtggtcatgg aaggttcggt caacggccac caattcaaat gcacaggtga aggagaaggc
     6121 aatccgtaca tgggaactca aaccatgagg atcaaagtca tcgagggagg acccctgcca
     6181 tttgcctttg acattcttgc cacgtcgttc atgtatggca gccgtacttt tatcaagtac
     6241 ccgaaaggca ttcctgattt ctttaaacag tcctttcctg agggttttac ttgggaaaga
     6301 gttacgagat acgaagatgg tggagtcgtc accgtcatgc aggacaccag ccttgaggat
     6361 ggctgtctcg tttaccacgt ccaagtcaga ggggtaaact ttccctccaa tggtcccgtg
     6421 atgcagaaaa agaccaaggg ttgggagcct aatacagaga tgatgtatcc agcagatggt
     6481 ggtctgaggg gatacactca tatggcactg aaagttgatg gtggtggcca tctgtcttgc
     6541 tctttcgtaa caacttacag gtcaaaaaag accgtcggga acatcaagat gcccggtatc
     6601 catgccgttg atcaccgcct ggaaaggtta gaggaaagtg acaatgaaat gttcgtagta
     6661 caacgcgaac acgcagttgc caagttcgcc gggcttggtg gtgggatgga cgagctgtac
     6721 aagggaagcg gagccactaa cttctccctg ttgaaacaag caggggatgt cgaagagaat
     6781 cccgggccaa tgactgccat ggaggagtca cagtcggata tcagcctcaa gagagcgctg
     6841 cccacctgca caagcgcctc tcccccgcaa aagaaaaaac cacttgatgg agagtatttc
     6901 accctcaaga tccgcgggcg taaacgcttc gagatgttcc gggagctgaa tgaggcctta
     6961 gagttaaagg atgcccatgc tacagaggag tctggagaca gcagggctca ctccagctac
     7021 ctgaagacca agaagggcca gtctacttcc cgccataaaa aaacaatggt caagaaagtg
     7081 gggcctgact cagactgata agaattccta gagctcgctg atcagcctcg actgtgcctt
     7141 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     7201 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     7261 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     7321 atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct
     7381 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     7441 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     7501 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     7561 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     7621 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     7681 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     7741 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     7801 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     7861 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     7921 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     7981 taacgcgaat tttaacaaaa ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     8041 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     8101 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     8161 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     8221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     8281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     8341 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     8401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     8461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     8521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     8581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     8641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     8701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     8761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     8821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     8881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     9001 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     9061 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     9121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     9181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     9241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     9301 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     9361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     9421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     9481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     9541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     9601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     9661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     9721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     9781 ttacggttcc tggccttttg ctggcctttt gctcacatgc tgggcccag
//
