LOCUS       sequence-329901-       11158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for a sgRNA against BFP and spCas9-Sa163 RT fusion
            protein linked to mCherry via a T2A peptide. Donor template An was 
            inserted in the Retron Sa163 msr-msd region by SpeI and AvrII..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11158)
  AUTHORS   Zhao B, Chen SA, Lee J, Fraser HB
  TITLE     Bacterial Retrons Enable Precise Gene Editing in Human Cells.
  JOURNAL   CRISPR J. 2022 Jan 24. doi: 10.1089/crispr.2021.0065.
  PUBMED    35076284
REFERENCE   2  (bases 1 to 11158)
  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: "CRISPR J. 
            2022 Jan 24. doi: 10.1089/crispr.2021.0065."
COMMENT     Sequence Label: 
            pBZ202-pU6-sgBFP-An_CBh-sv40NLS-Cas9-Sa163RT-NLS-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..11158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        782..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1069..1346
                     /label=chicken beta-actin promoter
     intron          1347..1575
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1587..1596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1596..1661
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1668..1688
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1713..5813
                     /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             1713..5813
                     /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             7353..7400
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7404..7457
                     /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             7464..8174
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7609..7627)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7877..7896
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(8199..8216)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8205..8412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8421..8561
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8421..8550
                     /label=AAV2 ITR
     rep_origin      8636..9091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8723..8742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8933..8954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9108..9127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9227..9249
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9287..9305)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9373..9477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9478..9546,9547..10338)
                     /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(9696..9715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10509..11097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10998..11017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ctgaagcact gcacgccatg ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgccgtac ggtgcgagcg
      361 accgagagag gtcccaagcc atcagcctca gcgcctcgag cgcgagagcg gcgttgcgcc
      421 gctctggttg aattgcagga cactctccgc aaggtagcct gttcttggct ctcttccctc
      481 cggtgagtac ctctccggcc ggggagctga accaacgaac tagtaagtcg tgctgcttca
      541 tgtggtcggg gtagcggctg aagcactgca cgccgtacgt cagggtggtc acgagggtgg
      601 gccagggcac gggcagcttg ccggtggtgc agatgaactt caggcctagg cgcaaccgcc
      661 gtttccccgg ccggagaggt actcaccgga ggggagagcc ggtgaggcta ccgtgcccca
      721 ggtgagaagg tggtgccttc gggcctccct cgaccgctcg cgcttttttt ctagaggtac
      781 ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      841 tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
     1081 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
     1141 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag
     1201 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
     1261 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     1321 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgac gctgccttcg ccccgtgccc
     1381 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1441 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg
     1501 tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa
     1561 atcacttttt ttcaggttgg accggtgcca ccatggacta taaggaccac gacggagact
     1621 acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca aagaagaagc
     1681 ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca
     1741 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga
     1801 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc
     1861 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa
     1921 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg
     1981 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata
     2041 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga
     2101 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc
     2161 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg
     2221 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga
     2281 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca
     2341 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg
     2401 gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca
     2461 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct
     2521 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc
     2581 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg
     2641 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg
     2701 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt
     2761 tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag
     2821 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg
     2881 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc
     2941 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc
     3001 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact
     3061 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg
     3121 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct
     3181 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc
     3241 acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga
     3301 ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc
     3361 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga
     3421 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc
     3481 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg
     3541 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga
     3601 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc
     3661 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc
     3721 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca
     3781 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag
     3841 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc
     3901 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga
     3961 tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc
     4021 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc
     4081 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc
     4141 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca
     4201 accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact
     4261 ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc
     4321 cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc
     4381 tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac
     4441 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg
     4501 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc
     4561 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc
     4621 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg
     4681 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg
     4741 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca
     4801 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta
     4861 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg
     4921 agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc
     4981 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc
     5041 tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt
     5101 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
     5161 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa
     5221 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
     5281 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
     5341 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
     5401 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
     5461 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
     5521 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
     5581 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
     5641 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
     5701 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga
     5761 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc gactctggag
     5821 gatctagcgg aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag
     5881 agagcagtgg cggcagcagc ggcggcagca gcatgacggc aaaactggaa agccacgtgc
     5941 ctgctgcacc accagtctca gcggaagcgc cggcaccgac gcgacctgac gctgctaagc
     6001 aggaggctcg acgagcccac catgaagccc tgagactgcg gtggaaagca atcgaggaag
     6061 ctgggggcac tgacgcctgg gtacgacagc agttggttgc gaaaggcgta gcggcggaag
     6121 aggtggactt cgaaagtctc tccgacaagc aaaaagcggc ttggaaagag aaaaaaaaag
     6181 ctgaggcaac cgagcggagg gctcaaaaac gacttgcctg ggaagcctgg aaagcgaccc
     6241 atattcatca tttgggcgtt ggagttcact gggacgaagc cggcgggcct gacaagttcg
     6301 atgtggcagg gagggaagaa agggcaaagg caaacgggtt gcccgaaggt cttgacagcg
     6361 tagaagcatt ggcgaaagcg ctgggaatct cagtatcccg attgcgctgg ttctcttttc
     6421 acagggaagt tgatacaggg acacactacc aaacttggga gatcccaaag agggatgggg
     6481 gcaaacgcac cctcacggcc ccgaaaaggg agcttaaggc agtacaaagg tgggtactcg
     6541 ccaatgtggt ggaaagactt cccgtacacg gagctgctca tgggtttgtt gcaggacgct
     6601 ccatactcac taatgcgctc gctcaccaag gagctgacgt ggttgttaaa gttgatatga
     6661 aagatttttt tcccagtgtc acatggcctc gagttaaagg cctgctcaga aagggaggtt
     6721 tgcctgaaaa cttggcgacc ctgctcgccc ttctgagtac cgaagctcca cgggaagtag
     6781 tccgattcag aggggaaacg ctgtacgtcg ctaagggccc acgcgctctg ccacagggcg
     6841 cgcctaccag tcctgcgctt actaacgcat tgtgtcttag gttggataag cggttgagcg
     6901 cgctgagtaa gcggttgggt ttcacataca ccagatacgc tgacgatctt acatttagtt
     6961 ggaggcgcgc taaaaaatct cgccagaaag agttgccgct ggctgatgct cccgtcgcgc
     7021 ttctcctggc ccgggttaag ggagtgttgg aagcagaagg tttcactttg catcctgata
     7081 agaccagggt tcagcggaag ggaagccggc aacgagtgac cggcctcgtg gttaatgaag
     7141 cgcccgaagg agtgccaggt gcccgagttc cccgagatgt tgttcggaga ttgcgagcgg
     7201 caatacacaa tagagagcaa ggtaaaccag ggccgactgg agaaacgttg gaacaactca
     7261 aaggcttggc agcttttctg cacatgacag acgctgaaaa aggaagagcg tttctgagaa
     7321 gacttgaggc cctggagaag aggcaaactg cgaaaaggcc ggcggccacg aaaaaggccg
     7381 gccaggcaaa aaagaaaaag cttgagggca gaggaagtct gctaacatgc ggtgacgtgg
     7441 aggagaatcc cggccctgct agcatggtga gcaagggcga ggaggataac atggccatca
     7501 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     7561 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     7621 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     7681 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     7741 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     7801 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     7861 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     7921 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     7981 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     8041 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     8101 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     8161 agctgtacaa gtgagaattc ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     8221 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     8281 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     8341 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag
     8401 gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     8461 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     8521 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     8581 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     8641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     8701 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     8761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     8821 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     8881 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     8941 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     9001 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     9061 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     9121 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     9181 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     9241 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     9301 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     9361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     9421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     9481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     9541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     9601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     9661 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     9721 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     9781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     9841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     9901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     9961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
    10021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
    10081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
    10141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
    10201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
    10261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
    10321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
    10381 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
    10441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    10501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    10561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    10621 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
    10681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    10741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    10801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    10861 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
    10921 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    10981 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    11041 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    11101 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
