LOCUS       sequence-251691-        9033 bp    DNA     circular SYN 14-APR-2025
DEFINITION  CMV promoter expression plasmid for bpNLS-TadA7.10(K20A/R21A)-32AA 
            linker-hSpCas9n(D10A)-bpNLS-P2A-EGFP-NLS (miniABEmax with K20A/R21A 
            mutations)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9033)
  AUTHORS   Grunewald J, Zhou R, Iyer S, Lareau CA, Garcia SP, Aryee MJ, Joung 
            JK
  TITLE     CRISPR DNA base editors with reduced RNA off-target and self-editing
            activities.
  JOURNAL   Nat Biotechnol. 2019 Sep 2. pii: 10.1038/s41587-019-0236-6. doi: 
            10.1038/s41587-019-0236-6.
  PUBMED    31477922
REFERENCE   2  (bases 1 to 9033)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2019 Sep 2. pii: 10.1038/s41587-019-0236-6. doi: 
            10.1038/s41587-019-0236-6."
COMMENT     Sequence Label: SECURE miniABEmax-K20A/R21A (pJUL1774)
FEATURES             Location/Qualifiers
     source          1..9033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    607..628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(643..648,649..666,667..673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    681..697
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     686..708
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     705..721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     705..721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    792..1016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1005..1022
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1045..1062)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1071..1091)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(1104..1814,1815..1817,1818..1820))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1104..1814,1815..1817,1818..1820))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1146..1167)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1495..1514
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(1821..1877)
                     /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             complement(1887..1907)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(1950..6023,6024..6026,6027..6050))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     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(1950..6050)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     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(6645..6665)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6714..6733)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6715..6733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6775..6978)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6804..6824)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6979..7358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7530..7549
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7628..7732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7733..7801,7802..8593)
                     /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(7951..7970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8764..9033,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      601 cgcaattaat gtgagttagc tcactcatta ggcaccctag gctttacact ttatgcttcc
      661 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      721 ccatgattac gccaagctct agctagaggt cgacggtatc gagccccagc tggttctttc
      781 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
      841 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
      901 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
      961 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     1021 ggctgatcag cgggtttaaa ctcaatggtg atggtgatga tgaccggtta gactttcctc
     1081 ttcttcttgg gagaaccacc agacttgtac agctcgtcca tgccgagagt gatcccggcg
     1141 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
     1201 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     1261 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
     1321 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     1381 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1441 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1501 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1561 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcactcca
     1621 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1681 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1741 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1801 tcctcgccct tgctcaccat aggtccaggg ttctcctcca cgtctccagc ctgcttcagc
     1861 aggctgaagt tagtagctcc gcttccgact ttcctcttct tcttgggctc gaattcgctg
     1921 ccgtcggcgg ttctttttga gccgccagag tcacctccca gctgagacag gtcgatccgt
     1981 gtctcgtaca ggccggtgat gctctggtgg atcagggtgg cgtccagcac ctctttggtg
     2041 ctggtgtacc tcttccggtc gatggtggtg tcaaagtact tgaaggcggc aggggctccc
     2101 agattggtca gggtaaacag gtggatgata ttctcggcct gctctctgat gggcttatcc
     2161 cggtgcttgt tgtaggcgga cagcactttg tccagattag cgtcggccag gatcactctc
     2221 ttggagaact cgctgatctg ctcgatgatc tcgtccaggt agtgcttgtg ctgttccaca
     2281 aacagctgtt tctgctcatt atcctcgggg gagcccttca gcttctcata gtggctggcc
     2341 aggtacagga agttcacata tttggagggc agggccagtt cgtttccctt ctgcagttcg
     2401 ccggcagagg ccagcattct cttccggccg ttttccagct cgaacaggga gtacttaggc
     2461 agcttgatga tcaggtcctt tttcacttct ttgtagccct tggcttccag aaagtcgatg
     2521 ggattcttct cgaagctgct tctttccatg atggtgatcc ccagcagctc tttcacactc
     2581 ttcagtttct tggacttgcc cttttccact ttggccacca ccagcacaga ataggccacg
     2641 gtggggctgt cgaagccgcc gtacttctta gggtcccagt ccttctttct ggcgatcagc
     2701 ttatcgctgt tcctcttggg caggatagac tctttgctga agccgcctgt ctgcacctcg
     2761 gtctttttca cgatattcac ttggggcatg ctcagcactt tccgcacggt ggcaaaatcc
     2821 cggcccttat cccacacgat ctccccggtt tcgccgtttg tctcgatcag aggccgcttc
     2881 cggatctcgc cgttggccag ggtaatctcg gtcttgaaaa agttcatgat gttgctgtag
     2941 aagaagtact tggcggtagc cttgccgatt tcctgctcgc tcttggcgat catcttccgc
     3001 acgtcgtaca ccttgtagtc gccgtacacg aactcgcttt ccagcttagg gtactttttg
     3061 atcagggcgg ttcccacgac ggcgttcagg taggcgtcgt gggcgtggtg gtagttgttg
     3121 atctcgcgca ctttgtaaaa ctggaaatcc ttccggaaat cggacaccag cttggacttc
     3181 agggtgatca ctttcacttc ccggatcagc ttgtcattct cgtcgtactt agtgttcatc
     3241 cgggagtcca ggatctgtgc cacgtgcttt gtgatctgcc gggtttccac cagctgtctc
     3301 ttgatgaagc cggccttatc cagttcgctc aggccgcctc tctcggcctt ggtcagattg
     3361 tcgaactttc tctgggtaat cagcttggcg ttcagcagct gccgccagta gttcttcatc
     3421 ttcttcacga cctcttcgga gggcacgttg tcgctcttgc cccggttctt gtcgcttctg
     3481 gtcagcacct tgttgtcgat ggagtcgtcc ttcagaaagc tctgaggcac gatatggtcc
     3541 acatcgtagt cggacagccg gttgatgtcc agttcctggt ccacgtacat atcccgccca
     3601 ttctgcaggt agtacaggta cagcttctcg ttctgcagct gggtgttttc cacggggtgt
     3661 tctttcagga tctggctgcc cagctctttg atgccctctt cgatccgctt cattctctcg
     3721 cggctgttct tctgtccctt ctgggtggtc tggttctctc tggccatttc gatcacgatg
     3781 ttctcgggct tgtgccggcc catcactttc acgagctcgt ccaccacctt cactgtctgc
     3841 aggatgccct tcttaatggc ggggctgccg gccagattgg caatgtgctc gtgcaggcta
     3901 tcgccctggc cggacacctg ggctttctgg atgtcctctt taaaggtcag gctgtcgtcg
     3961 tggatcagct gcatgaagtt tctgttggcg aagccgtcgg acttcaggaa atccaggatt
     4021 gtcttgccgg actgcttgtc ccggatgccg ttgatcagct tccggctcag cctgccccag
     4081 ccggtgtatc tccgccgctt cagctgcttc atcactttgt cgtcgaacag gtgggcatag
     4141 gttttcagcc gttcctcgat catctctctg tcctcaaaca gtgtcagggt cagcacgata
     4201 tcttccagaa tgtcctcgtt ttcctcattg tccaggaagt ccttgtcctt gataattttc
     4261 agcagatcgt ggtatgtgcc cagggaggcg ttgaaccgat cttccacgcc ggagatttcc
     4321 acggagtcga agcactcgat tttcttgaag tagtcctctt tcagctgctt cacggtcact
     4381 ttccggttgg tcttgaacag caggtccacg atggcctttt tctgctcgcc gctcaggaag
     4441 gcgggctttc tcattccctc ggtcacgtat ttcactttgg tcagctcgtt atacacggtg
     4501 aagtactcgt acagcaggct gtgcttgggc agcaccttct cgttgggcag gttcttatcg
     4561 aagttggtca tccgctcgat gaagctctgg gcggaagcgc ccttgtccac cacttcctcg
     4621 aagttccagg gggtgatggt ttcctcgctc tttctggtca tccaggcgaa tctgctgttt
     4681 cccctggcca gagggcccac gtagtagggg atgcggaagg tcaggatctt ctcgatcttt
     4741 tcccggttgt ccttcaggaa tgggtaaaaa tcttcctgcc gccgcagaat ggcgtgcagc
     4801 tctcccaggt ggatctggtg ggggatgctg ccgttgtcga aggtccgctg cttccgcagc
     4861 aggtcctctc tgttcagctt cacgagcagt tcctcggtgc cgtccatctt ttccaggatg
     4921 ggcttgatga acttgtagaa ctcttcctgg ctggctccgc cgtcaatgta gccggcgtag
     4981 ccgttcttgc tctggtcgaa gaaaatctct ttgtacttct caggcagctg ctgccgcacg
     5041 agagctttca gcagggtcag gtcctggtgg tgctcgtcgt atctcttgat catagaggcg
     5101 ctcagggggg ccttggtgat ctcggtgttc actctcagga tgtcgctcag caggatggcg
     5161 tcggacaggt tcttggcggc cagaaacagg tcggcgtact ggtcgccgat ctgggccagc
     5221 aggttgtcca ggtcgtcgtc gtaggtgtcc ttgctcagct gcagtttggc atcctcggcc
     5281 aggtcgaagt tgctcttgaa gttgggggtc aggcccaggc tcagggcaat caggtttccg
     5341 aacaggccat tcttcttctc gccgggcagc tgggcgatca gattttccag ccgtctgctc
     5401 ttgctcagtc tggcagacag gatggccttg gcgtccacgc cgctggcgtt gatggggttt
     5461 tcctcgaaca gctggttgta ggtctgcacc agctggatga acagcttgtc cacgtcgctg
     5521 ttgtcggggt tcaggtcgcc ctcgatcagg aagtggcccc ggaacttgat catgtgggcc
     5581 agggccagat agatcagccg caggtcggcc ttgtcggtgc tgtccaccag tttctttctc
     5641 aggtggtaga tggtggggta cttctcgtgg taggccacct cgtccacgat gttgccgaag
     5701 atggggtgcc gctcgtgctt cttatcctct tccaccagga aggactcttc cagtctgtgg
     5761 aagaagctgt cgtccacctt ggccatctcg ttgctgaaga tctcttgcag atagcagatc
     5821 cggttcttcc gtctggtgta tcttcttctg gcggttctct tcagccgggt ggcctcggct
     5881 gtttcgccgc tgtcgaacag cagggctccg atcaggttct tcttgatgct gtgccggtcg
     5941 gtgttgccca gcaccttgaa tttcttgctg ggcaccttgt actcgtcggt gatcacggcc
     6001 cagcccacag agttggtgcc gatggccagg ccgatgctgt acttcttgtc tgaccccccg
     6061 ctgctgcccc cgctgctttc aggtgttgcg ctctcgcttg tgccaggtgt ctcagagcca
     6121 gaggagcctc cgctagatcc tccggagtcg gtggagctct gggccttctt ctgagcattg
     6181 aacacctgtc taggcatccg aaagaaatag cacagcaggg cggcacattc atctgccagg
     6241 attccctcgg taatttcgac gcggtgattc atgccggggt agtgcagcac gtccatcagg
     6301 gagcctgcgg cgccggtttt tgcgttcctc acgccaaaca ccacgcggcc gatcctagag
     6361 tggatcatgg cgccggcgca catcacgcaa ggctcgaatg tcacgtacag ggtggcgtca
     6421 atcagtctgt agttctgcat gaccaggccg ccctgtctca gggccataat ttcggcatgg
     6481 gctgttgggt cgtgcaggcc gatggctctg ttccagccct cgccgatcac tctattgttc
     6541 agcaccagca cggctcccac aggcacctcc ctctcatcgc gtgctgcagc ggccagggtc
     6601 agggcatgtc tcatccagta ctcgtgggaa aactccacct cagagacttt ccgcttcttc
     6661 tttggtgact cgaactcgct tccgtcggct gtccgtttca tggtggcggc tctccctata
     6721 gtgagtcgta ttagcggccg cggatctcta gcggatctga cggttcacta aaccagctct
     6781 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     6841 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     6901 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     6961 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     7021 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     7081 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     7141 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     7201 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     7261 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     7321 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     7381 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     7441 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     7501 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtcga
     7561 ccctagggga tcgggagatc gatctcccga tccgtcgacg tcaggtggca cttttcgggg
     7621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     8281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     8341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     8461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8701 cccttaacgt gagttttcgt tccactgagt gtcagacccc gtagaaaaga tcaaaggatc
     8761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9001 tgctgccagt ggcgataagt cgtgtcttac cgg
//
