LOCUS       sequence-342530-       11128 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 11128)
  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 11128)
  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..11128
                     /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"
     intron          1318..1545
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1557..1566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1566..1631
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1638..1658
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1683..5783
                     /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             1683..5783
                     /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             7323..7370
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7374..7427
                     /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             7434..8144
                     /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(7579..7597)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7847..7866
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(8169..8186)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8175..8382
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8391..8531
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8391..8520
                     /label=AAV2 ITR
     rep_origin      8606..9061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8693..8712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8903..8924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9078..9097)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9197..9219
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9257..9275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9343..9447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9448..9516,9517..10308)
                     /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(9666..9685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10479..11067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10968..10987
                     /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 gggggggcga ggggcggggc
     1201 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     1261 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     1321 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     1381 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     1441 ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta
     1501 atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca
     1561 ccatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
     1621 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
     1681 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     1741 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     1801 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     1861 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     1921 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     1981 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2041 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2101 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     2161 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2221 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     2281 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     2341 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     2401 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     2461 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     2521 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     2581 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     2641 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     2701 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     2761 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     2821 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     2881 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     2941 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3001 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3061 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3121 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     3181 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3241 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     3301 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     3361 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     3421 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     3481 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     3541 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     3601 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     3661 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     3721 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     3781 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     3841 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     3901 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     3961 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     4021 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     4081 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     4141 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     4201 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     4261 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     4321 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     4381 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     4441 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     4501 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     4561 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     4621 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     4681 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     4741 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     4801 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     4861 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     4921 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     4981 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     5041 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     5101 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     5161 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     5221 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     5281 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     5341 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     5401 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     5461 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     5521 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     5581 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     5641 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     5701 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     5761 acctgtctca gctgggaggc gactctggag gatctagcgg aggatcctct ggcagcgaga
     5821 caccaggaac aagcgagtca gcaacaccag agagcagtgg cggcagcagc ggcggcagca
     5881 gcatgacggc aaaactggaa agccacgtgc ctgctgcacc accagtctca gcggaagcgc
     5941 cggcaccgac gcgacctgac gctgctaagc aggaggctcg acgagcccac catgaagccc
     6001 tgagactgcg gtggaaagca atcgaggaag ctgggggcac tgacgcctgg gtacgacagc
     6061 agttggttgc gaaaggcgta gcggcggaag aggtggactt cgaaagtctc tccgacaagc
     6121 aaaaagcggc ttggaaagag aaaaaaaaag ctgaggcaac cgagcggagg gctcaaaaac
     6181 gacttgcctg ggaagcctgg aaagcgaccc atattcatca tttgggcgtt ggagttcact
     6241 gggacgaagc cggcgggcct gacaagttcg atgtggcagg gagggaagaa agggcaaagg
     6301 caaacgggtt gcccgaaggt cttgacagcg tagaagcatt ggcgaaagcg ctgggaatct
     6361 cagtatcccg attgcgctgg ttctcttttc acagggaagt tgatacaggg acacactacc
     6421 aaacttggga gatcccaaag agggatgggg gcaaacgcac cctcacggcc ccgaaaaggg
     6481 agcttaaggc agtacaaagg tgggtactcg ccaatgtggt ggaaagactt cccgtacacg
     6541 gagctgctca tgggtttgtt gcaggacgct ccatactcac taatgcgctc gctcaccaag
     6601 gagctgacgt ggttgttaaa gttgatatga aagatttttt tcccagtgtc acatggcctc
     6661 gagttaaagg cctgctcaga aagggaggtt tgcctgaaaa cttggcgacc ctgctcgccc
     6721 ttctgagtac cgaagctcca cgggaagtag tccgattcag aggggaaacg ctgtacgtcg
     6781 ctaagggccc acgcgctctg ccacagggcg cgcctaccag tcctgcgctt actaacgcat
     6841 tgtgtcttag gttggataag cggttgagcg cgctgagtaa gcggttgggt ttcacataca
     6901 ccagatacgc tgacgatctt acatttagtt ggaggcgcgc taaaaaatct cgccagaaag
     6961 agttgccgct ggctgatgct cccgtcgcgc ttctcctggc ccgggttaag ggagtgttgg
     7021 aagcagaagg tttcactttg catcctgata agaccagggt tcagcggaag ggaagccggc
     7081 aacgagtgac cggcctcgtg gttaatgaag cgcccgaagg agtgccaggt gcccgagttc
     7141 cccgagatgt tgttcggaga ttgcgagcgg caatacacaa tagagagcaa ggtaaaccag
     7201 ggccgactgg agaaacgttg gaacaactca aaggcttggc agcttttctg cacatgacag
     7261 acgctgaaaa aggaagagcg tttctgagaa gacttgaggc cctggagaag aggcaaactg
     7321 cgaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag cttgagggca
     7381 gaggaagtct gctaacatgc ggtgacgtgg aggagaatcc cggccctgct agcatggtga
     7441 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     7501 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     7561 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     7621 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     7681 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     7741 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     7801 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     7861 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     7921 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     7981 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     8041 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     8101 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtgagaattc ctagagctcg
     8161 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     8221 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     8281 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     8341 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
     8401 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     8461 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     8521 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     8581 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8641 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     8701 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8761 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8821 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8881 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     8941 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     9001 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     9061 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     9121 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     9181 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     9241 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     9301 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     9361 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     9421 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     9481 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     9541 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     9601 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     9661 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     9721 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     9781 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     9841 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     9901 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     9961 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    10021 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    10081 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    10141 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
    10201 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    10261 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    10321 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    10381 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    10441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
    10501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
    10561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
    10621 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    10681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    10741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    10801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    10861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    10921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    10981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    11041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
    11101 gccttttgct ggccttttgc tcacatgt
//
