LOCUS       sequence-476973-       11471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression SpCas9 and Efe1-RT from bidirectional CAG promoters. 
            SpCas9 sgRNA and Efe1 non-coding expression is driven by U6 and H1 
            respecitvely..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11471)
  AUTHORS   Buffington JD, Kuo H, Hu K, Chang Y, Javanmardi K, Voigt B, Li Y, 
            Little ME, Devanathan SK, Xhemalce B, Gray RS, Finkelstein IJ
  TITLE     Discovery and Engineering of Retrons for Precise Genome Editing
  JOURNAL   bioRxiv 2024.07.21.604473; doi: 10.1101/2024.07.21.604473
REFERENCE   2  (bases 1 to 11471)
  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: "bioRxiv 
            2024.07.21.604473; doi: 10.1101/2024.07.21.604473"
COMMENT     Sequence Label: pIF1079
FEATURES             Location/Qualifiers
     source          1..11471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    690..897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     886..903
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(931..978)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(979..5079)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(979..5079)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(5104..5124)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5131..5196)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      5196..5205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          5217..5444
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(5445..5727)
                     /label=chicken beta-actin promoter
     enhancer        5729..6014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     primer_bind     complement(6334..6353)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        complement(6506..6581)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(6615..6865)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6675..6694)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6845..6865)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     enhancer        7324..7609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        7611..7891
                     /label=chicken beta-actin promoter
     intron          7892..8119
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             8140..8160
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9115..9162
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    9196..9403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9600..10055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9687..9706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9897..9918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10072..10091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10250..10268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10336..10440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10441..10509,10510..11301)
                     /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(10659..10678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tttagtctat
      661 cctcaatgtc taggggttcc tgcggccgct ccccagcatg cctgctattc tcttcccaat
      721 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
      781 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
      841 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
      901 aggctctcta ggaattcgtc ttcctcatca ctttttcttt tttgcctggc cggccttttt
      961 cgtggccgcc ggccttttgt cgcctcccag ctgagacagg tcgatccgtg tctcgtacag
     1021 gccggtgatg ctctggtgga tcagggtggc gtccagcacc tctttggtgc tggtgtacct
     1081 cttccggtcg atggtggtgt caaagtactt gaaggcggca ggggctccca gattggtcag
     1141 ggtaaacagg tggatgatat tctcggcctg ctctctgatg ggcttatccc ggtgcttgtt
     1201 gtaggcggac agcactttgt ccagattagc gtcggccagg atcactctct tggagaactc
     1261 gctgatctgc tcgatgatct cgtccaggta gtgcttgtgc tgttccacaa acagctgttt
     1321 ctgctcatta tcctcggggg agcccttcag cttctcatag tggctggcca ggtacaggaa
     1381 gttcacatat ttggagggca gggccagttc gtttcccttc tgcagttcgc cggcagaggc
     1441 cagcattctc ttccggccgt tttccagctc gaacagggag tacttaggca gcttgatgat
     1501 caggtccttt ttcacttctt tgtagccctt ggcttccaga aagtcgatgg gattcttctc
     1561 gaagctgctt ctttccatga tggtgatccc cagcagctct ttcacactct tcagtttctt
     1621 ggacttgccc ttttccactt tggccaccac cagcacagaa taggccacgg tggggctgtc
     1681 gaagccgccg tacttcttag ggtcccagtc cttctttctg gcgatcagct tatcgctgtt
     1741 cctcttgggc aggatagact ctttgctgaa gccgcctgtc tgcacctcgg tctttttcac
     1801 gatattcact tggggcatgc tcagcacttt ccgcacggtg gcaaaatccc ggcccttatc
     1861 ccacacgatc tccccggttt cgccgtttgt ctcgatcaga ggccgcttcc ggatctcgcc
     1921 gttggccagg gtaatctcgg tcttgaaaaa gttcatgatg ttgctgtaga agaagtactt
     1981 ggcggtagcc ttgccgattt cctgctcgct cttggcgatc atcttccgca cgtcgtacac
     2041 cttgtagtcg ccgtacacga actcgctttc cagcttaggg tactttttga tcagggcggt
     2101 tcccacgacg gcgttcaggt aggcgtcgtg ggcgtggtgg tagttgttga tctcgcgcac
     2161 tttgtaaaac tggaaatcct tccggaaatc ggacaccagc ttggacttca gggtgatcac
     2221 tttcacttcc cggatcagct tgtcattctc gtcgtactta gtgttcatcc gggagtccag
     2281 gatctgtgcc acgtgctttg tgatctgccg ggtttccacc agctgtctct tgatgaagcc
     2341 ggccttatcc agttcgctca ggccgcctct ctcggccttg gtcagattgt cgaactttct
     2401 ctgggtaatc agcttggcgt tcagcagctg ccgccagtag ttcttcatct tcttcacgac
     2461 ctcttcggag ggcacgttgt cgctcttgcc ccggttcttg tcgcttctgg tcagcacctt
     2521 gttgtcgatg gagtcgtcct tcagaaagct ctgaggcacg atatggtcca catcgtagtc
     2581 ggacagccgg ttgatgtcca gttcctggtc cacgtacata tcccgcccat tctgcaggta
     2641 gtacaggtac agcttctcgt tctgcagctg ggtgttttcc acggggtgtt ctttcaggat
     2701 ctggctgccc agctctttga tgccctcttc gatccgcttc attctctcgc ggctgttctt
     2761 ctgtcccttc tgggtggtct ggttctctct ggccatttcg atcacgatgt tctcgggctt
     2821 gtgccggccc atcactttca cgagctcgtc caccaccttc actgtctgca ggatgccctt
     2881 cttaatggcg gggctgccgg ccagattggc aatgtgctcg tgcaggctat cgccctggcc
     2941 ggacacctgg gctttctgga tgtcctcttt aaaggtcagg ctgtcgtcgt ggatcagctg
     3001 catgaagttt ctgttggcga agccgtcgga cttcaggaaa tccaggattg tcttgccgga
     3061 ctgcttgtcc cggatgccgt tgatcagctt ccggctcagc ctgccccagc cggtgtatct
     3121 ccgccgcttc agctgcttca tcactttgtc gtcgaacagg tgggcatagg ttttcagccg
     3181 ttcctcgatc atctctctgt cctcaaacag tgtcagggtc agcacgatat cttccagaat
     3241 gtcctcgttt tcctcattgt ccaggaagtc cttgtccttg ataattttca gcagatcgtg
     3301 gtatgtgccc agggaggcgt tgaaccgatc ttccacgccg gagatttcca cggagtcgaa
     3361 gcactcgatt ttcttgaagt agtcctcttt cagctgcttc acggtcactt tccggttggt
     3421 cttgaacagc aggtccacga tggccttttt ctgctcgccg ctcaggaagg cgggctttct
     3481 cattccctcg gtcacgtatt tcactttggt cagctcgtta tacacggtga agtactcgta
     3541 cagcaggctg tgcttgggca gcaccttctc gttgggcagg ttcttatcga agttggtcat
     3601 ccgctcgatg aagctctggg cggaagcgcc cttgtccacc acttcctcga agttccaggg
     3661 ggtgatggtt tcctcgctct ttctggtcat ccaggcgaat ctgctgtttc ccctggccag
     3721 agggcccacg tagtagggga tgcggaaggt caggatcttc tcgatctttt cccggttgtc
     3781 cttcaggaat gggtaaaaat cttcctgccg ccgcagaatg gcgtgcagct ctcccaggtg
     3841 gatctggtgg gggatgctgc cgttgtcgaa ggtccgctgc ttccgcagca ggtcctctct
     3901 gttcagcttc acgagcagtt cctcggtgcc gtccatcttt tccaggatgg gcttgatgaa
     3961 cttgtagaac tcttcctggc tggctccgcc gtcaatgtag ccggcgtagc cgttcttgct
     4021 ctggtcgaag aaaatctctt tgtacttctc aggcagctgc tgccgcacga gagctttcag
     4081 cagggtcagg tcctggtggt gctcgtcgta tctcttgatc atagaggcgc tcaggggggc
     4141 cttggtgatc tcggtgttca ctctcaggat gtcgctcagc aggatggcgt cggacaggtt
     4201 cttggcggcc agaaacaggt cggcgtactg gtcgccgatc tgggccagca ggttgtccag
     4261 gtcgtcgtcg taggtgtcct tgctcagctg cagtttggca tcctcggcca ggtcgaagtt
     4321 gctcttgaag ttgggggtca ggcccaggct cagggcaatc aggtttccga acaggccatt
     4381 cttcttctcg ccgggcagct gggcgatcag attttccagc cgtctgctct tgctcagtct
     4441 ggcagacagg atggccttgg cgtccacgcc gctggcgttg atggggtttt cctcgaacag
     4501 ctggttgtag gtctgcacca gctggatgaa cagcttgtcc acgtcgctgt tgtcggggtt
     4561 caggtcgccc tcgatcagga agtggccccg gaacttgatc atgtgggcca gggccagata
     4621 gatcagccgc aggtcggcct tgtcggtgct gtccaccagt ttctttctca ggtggtagat
     4681 ggtggggtac ttctcgtggt aggccacctc gtccacgatg ttgccgaaga tggggtgccg
     4741 ctcgtgcttc ttatcctctt ccaccaggaa ggactcttcc agtctgtgga agaagctgtc
     4801 gtccaccttg gccatctcgt tgctgaagat ctcttgcaga tagcagatcc ggttcttccg
     4861 tctggtgtat cttcttctgg cggttctctt cagccgggtg gcctcggctg tttcgccgct
     4921 gtcgaacagc agggctccga tcaggttctt cttgatgctg tgccggtcgg tgttgcccag
     4981 caccttgaat ttcttgctgg gcaccttgta ctcgtcggtg atcacggccc agcccacaga
     5041 gttggtgccg atgtccaggc cgatgctgta cttcttgtcg gctgctggga ctccgtggat
     5101 accgaccttc cgcttcttct ttggggccat cttatcgtca tcgtctttgt aatcaatatc
     5161 atgatccttg tagtctccgt cgtggtcctt atagtccatg gtggcagaag acggaactga
     5221 aaaaaagtga tttcaggcag gtgctccagg taattaaaca ttaatacccc accaaccaac
     5281 catcccttaa acccttacct cttgctcagc taattacagc ccggaggaga agggccgtcc
     5341 cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg
     5401 gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg ccgcgcgctt
     5461 cgctttttat agggccgccg ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg
     5521 ccgctctgat tggctgccgc cgcacctctc cgcctcgccc cgccccgccc ctcgccccgc
     5581 cccgccccgc ctggcgcgcg cccccccccc cccccccccg cccccatcgc tgcacaaaat
     5641 aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggggag atggggagag
     5701 tgaagcagaa cgtggggctc acctcgacca tggtaatagc gatgactaat acgtagatgt
     5761 actgccaagt aggaaagtcc cataaggtca tgtactgggc acaatgccag gcgggccatt
     5821 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     5881 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     5941 ttactattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     6001 taagttatgt aacgcgagac ctgctacgta acgggagggt cgctagaaaa aaacgccaac
     6061 agcggcaata ccatcaaacc attacaaacg gcagaagctg gaggaggaag ggcctgagtc
     6121 cgagcagaag aaagcatgta gcgaagggct cccatcacat caaccggtgg cgcattgcca
     6181 cgaagcaggc caactggcgt taagtcggcg actcgccgac cctcccggca caaaaggccg
     6241 gaaacgctat tgccactgct ggcgggggat ctgtggtcct catacagaac ttataagatt
     6301 cccaaatcca aagacatttc acgtttatgg tgatttccca gaacacatag cgacatgcaa
     6361 atattgcagg gcgccactcc cctgtccctc acagccatct tcctgccagg gcgcacgcgc
     6421 gctgggtgtt cccgcctagt gacactgggc ccgcgattcc ttggagcggg ttgatgacgt
     6481 cagcgttccg tacgctagaa aaaaagcacc gactcggtgc cactttttca agttgataac
     6541 ggactagcct tattttaact tgctatttct agctctaaaa cttcttcttc tgctcggact
     6601 cggtgtttcg tcctggtgtt taggtctcct tccacaagat atataaagcc aagaaatcga
     6661 aatactttca agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg
     6721 caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt
     6781 ttacagtcaa attaattcca attatctctc taacagcctt gtatcgtata tgcaaatatg
     6841 aaggaatcat gggaaatagg ccctcacatt acggggtctg acgctcagtg gaacgaaaac
     6901 tcacgttaag ggattttggt catgagatta tcaaaaagga tggcgacgaa tagcagttta
     6961 cttcctccct gcgggcccct cctgaggtgc cacctgtacg gctcgttgaa gccgattagt
     7021 acaatagatt tattcaaccc caaaggtcta cactcccggc ctactcttag ctgatatgtc
     7081 gcgtcgggcc ccgggtttga gtgcgttagc gccgggttgt ctggcccagt tcgacttctg
     7141 ggtacctcat cgggttacac tggctgacac tcatccttca gacagggttc ccttattggg
     7201 actagtaacc gaggttcgaa actatcccta cttccaatgg tctgcaccta gatcctttta
     7261 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7321 agccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     7381 attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7441 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     7501 tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga ccttatggga
     7561 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     7621 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     7681 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg gggggcgcgc
     7741 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     7801 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     7861 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
     7921 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     7981 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta
     8041 agggtttaag ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc
     8101 tgaaatcact ttttttcagg ttggaagagc gccaccatgc ctaagaaaaa gagaaaggtg
     8161 gacgccacca gaatcacact gctggccctt gacctgtttg gcagccctgg ctggtccgcc
     8221 gataaggaaa tccagaggct gtatgccctg agcaatcaca ccggcagaca ctaccggaga
     8281 atcatcctga gcaagaggca tggcggccag agactggtgc tggctcctga ttacctgctg
     8341 aaaaccgtgc agcggaacat tctgaaaaac gtgctgtccc agttccccct gtctagcttc
     8401 gccacagctt acagacctgg atgtccaatc gtgtctaatg cccagcccca ctgtcagcag
     8461 cctcagatcc tgaagttcga catcgagaac ttcttcgaca gcatctcctg gctgcaagtg
     8521 tggcgggtgt tccggcaggc tcagctgccc agaaacgtgg tcacaatgct gacctggctg
     8581 tgctgctaca acgacgccct gcctcaaggc gccccaacaa gccccgccat ctctaatctg
     8641 gtgatgtgca gattcgatga gagaatcgga gaatggtgcc aagctcgggg catcacctac
     8701 acccgctact gcgacgacat gaccttcagc ggccacttca acgccagact ggttaagaac
     8761 aaggtgtgcg gcctgctcgc cgagctgggc ctgaacctga accagagaaa gagctgcctg
     8821 gtggccgcct gcaagcggca acaggtgacc ggcatcgtgg tgaaccacaa gcctcagctg
     8881 gcccgggaag ccagacgggc cctgagacag gaggtgcacc tctgtcagaa atacggcgtg
     8941 atcagccacc tgagccacag aggtgagctg gacagctctg gagatctgca cgcccaggcc
     9001 accgcctacc tgtacgccct gcagggcaga attaactggc tgctgcagat caaccccgag
     9061 gacgagggct ttaagcaggc cagagaaggc gtgaagcgga tgctggtcgc ctggaagcgg
     9121 cctgctgcta caaagaaagc cggccaggcc aagaagaaga aatgagaatt ccgctcttct
     9181 agagctcgct gatcactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     9241 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     9301 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     9361 gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg caggaacccc
     9421 tagtgatgga gttggccact ccctctcggc gaccaaaggt cgcccgacgc ccgggctttg
     9481 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     9541 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     9601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     9661 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     9721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     9781 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     9841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     9901 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     9961 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
    10021 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
    10081 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
    10141 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacattct ccggagctgc
    10201 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggttaaaggg cctcgtgata
    10261 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
    10321 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
    10381 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
    10441 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
    10501 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
    10561 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
    10621 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
    10681 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
    10741 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
    10801 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
    10861 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
    10921 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
    10981 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
    11041 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
    11101 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
    11161 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
    11221 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
    11281 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
    11341 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
    11401 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
    11461 aaaggatctt c
//
