LOCUS       sequence-318808-       11101 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11101)
  AUTHORS   Zhang X, Zhu B, Chen L, Xie L, Yu W, Wang Y, Li L, Yin S, Yang L, Hu
            H, Han H, Li Y, Wang L, Chen G, Ma X, Geng H, Huang W, Pang X, Yang 
            Z, Wu Y, Siwko S, Kurita R, Nakamura Y, Yang L, Liu M, Li D
  TITLE     Dual base editor catalyzes both cytosine and adenine base 
            conversions in human cells.
  JOURNAL   Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0527-y. doi: 
            10.1038/s41587-020-0527-y.
  PUBMED    32483363
REFERENCE   2  (bases 1 to 11101)
  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. 2020 Jun 1. pii: 10.1038/s41587-020-0527-y. doi: 
            10.1038/s41587-020-0527-y."
COMMENT     Sequence Label: A&C-BEmax
FEATURES             Location/Qualifiers
     source          1..11101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..412
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     164..184
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     335..354
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        431..506
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        526..905
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        906..1109
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1060..1080
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1191..1211
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2988..3011,3012..3014,3015..7088)
                     /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             2988..7088
                     /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             7119..7367
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7398..7646
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7689..7709
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7719..7775
                     /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             join(7776..7778,7779..7781,7782..8492)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7776..7778,7779..7781,7782..8492)
                     /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(8082..8101)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8429..8450
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8505..8525
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8548..8565)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8554..8778
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8862..8884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8873..8889
                     /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)."
     promoter        complement(join(8897..8903,8904..8921,8922..8927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8942..8963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9080..9097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9251..9839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9331..9350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10010..10801,10802..10870))
                     /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     10633..10652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10871..10975)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11054..11073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
       61 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
      121 tgtacgggcc agatatacgc gtggatccct gcagttaatt aaagagggcc tatttcccat
      181 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
      241 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
      301 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
      361 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc
      421 gagaagacct gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      481 ttgaaaaagt ggcaccgagt cggtgctttt tttatgcatg gtaccgacat tgattattga
      541 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      601 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      661 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      721 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      781 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      841 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      901 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      961 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1021 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1081 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagagat
     1141 ccgcggccgc caccatgaag aggaccgccg atggctctga gttcgagagc cccaagaaga
     1201 agcggaaggt ggacagcctg ctgatgaacc ggagaaagtt cctgtatcag tttaagaatg
     1261 tgaggtgggc aaagggcagg cgcgagacct acctgtgcta tgtggtgaag cggagagata
     1321 gcgccacatc tttcagcctg gactttggct acctgcggaa caagaatggc tgccacgtgg
     1381 agctgctgtt tctgagatac atctccgact gggatctgga cccaggcagg tgttatcgcg
     1441 tgacctggtt cacatcctgg tctccctgct acgattgtgc ccggcacgtg gccgactttc
     1501 tgagaggcaa ccctaatctg tctctgagga tcttcaccgc ccgcctgtat ttttgcgagg
     1561 ataggaaggc cgagccagag ggactgaggc gcctgcacag ggccggcgtg cagatcgcca
     1621 tcatgacatt caaggactac ttttattgtt ggaacacctt cgtggagaat cacgagcgga
     1681 catttaaggc ctgggaggga ctgcacgaga actccgtgcg gctgtctaga cagctgcgga
     1741 gaatcctgct gccagaggcc gccgccaagg aagctgccgc caaggaggcc gccgccaaga
     1801 gcgaggtgga gttcagccac gagtactgga tgagacacgc cctgaccctg gctaagagag
     1861 cttgggacga gagagaggtc ccagtgggag ctgttctggt tcataacaac agggtgatcg
     1921 gcgagggatg gaacagacct atcgggagac acgacccaac cgctcatgct gaaatcatgg
     1981 ccctgagaca aggagggctg gtgatgcaaa attacagact gatcgacgca accctgtacg
     2041 tgaccctgga gccttgtgtg atgtgcgcag gagcaatgat ccactccaga atcggcagag
     2101 tggtgttcgg agctagagat gccaaaaccg gagccgctgg aagcctgatg gacgttctgc
     2161 atcaccccgg aatgaatcac agagtggaga taaccgaggg cattctggcc gacgagtgtg
     2221 ctgctctgct gtctgatttc ttcagaatga gaaggcagga aatcaaggcc cagaaaaagg
     2281 cccaaagcag caccgacagc ggaggatcta gcggaggatc aagcggaagc gagactcctg
     2341 gaaccagcga aagcgcaacc ccagaaagca gcggaggaag tagcggagga agctcagaag
     2401 tcgagttcag ccatgagtat tggatgagac atgctctgac cctggcaaag agagcaagag
     2461 acgagagaga ggtcccagtg ggagcagttc tggtgctgaa caacagagtg atcggggagg
     2521 ggtggaacag agcaatcgga ctgcacgacc ctacagcaca cgcagaaata atggcactga
     2581 gacaaggggg gctcgtgatg caaaactaca ggctgatcga cgccaccctg tacgtcacat
     2641 ttgagccctg tgtgatgtgt gccggagcca tgattcacag tagaatcggc cgggtggtgt
     2701 tcggtgtgag aaacgctaaa acaggcgccg ccggaagcct gatggatgtt ctgcattacc
     2761 ccggcatgaa tcaccgggtg gagatcacag agggcatcct ggctgacgaa tgtgccgctc
     2821 tgctgtgtta cttcttcaga atgccccgac aagtgttcaa cgcccagaag aaagcccagt
     2881 caagcaccga ctctggcgga tctagcggtg gatctagcgg ctctgagacc cctggaacat
     2941 ccgaatccgc cactccagag agcagcggag gctcttctgg aggatcagac aagaagtaca
     3001 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     3061 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     3121 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     3181 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     3241 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     3301 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     3361 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     3421 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     3481 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     3541 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     3601 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     3661 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     3721 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     3781 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     3841 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     3901 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     3961 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     4021 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     4081 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     4141 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     4201 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     4261 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     4321 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     4381 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     4441 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     4501 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     4561 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     4621 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     4681 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     4741 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     4801 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     4861 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     4921 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     4981 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     5041 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     5101 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     5161 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     5221 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     5281 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     5341 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     5401 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     5461 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     5521 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     5581 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     5641 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     5701 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     5761 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     5821 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     5881 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     5941 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     6001 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     6061 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     6121 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     6181 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     6241 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     6301 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     6361 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     6421 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     6481 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     6541 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     6601 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     6661 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     6721 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     6781 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     6841 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     6901 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     6961 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     7021 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     7081 gaggtgacag cggcgggagc ggcgggagcg gggggagcac taatctgagc gacatcattg
     7141 agaaggagac tgggaaacag ctggtcattc aggagtccat cctgatgctg cctgaggagg
     7201 tggaggaagt gatcggcaac aagccagagt ctgacatcct ggtgcacacc gcctacgacg
     7261 agtccacaga tgagaatgtg atgctgctga cctctgacgc ccccgagtat aagccttggg
     7321 ccctggtcat ccaggattct aacggcgaga ataagatcaa gatgctgagc ggaggatccg
     7381 gaggatctgg aggcagcacc aacctgtctg acatcatcga gaaggagaca ggcaagcagc
     7441 tggtcatcca ggagagcatc ctgatgctgc ccgaagaagt cgaagaagtg atcggaaaca
     7501 agcctgagag cgatatcctg gtccataccg cctacgacga gagtaccgac gaaaatgtga
     7561 tgctgctgac atccgacgcc ccagagtata agccctgggc tctggtcatc caggattcca
     7621 acggagagaa caaaatcaaa atgctgtctg gcggctcaaa aagaaccgcc gacggcagcg
     7681 aattcgagcc caagaagaag aggaaagtcg gaagcggagc tactaacttc agcctgctga
     7741 agcaggctgg agacgtggag gagaaccctg gacctatggt gagcaagggc gaggagctgt
     7801 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     7861 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     7921 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctatggag
     7981 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     8041 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     8101 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     8161 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     8221 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     8281 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     8341 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     8401 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     8461 ggatcactct cggcatggac gagctgtaca agtctggtgg ttctcccaag aagaaaagga
     8521 aagtctaagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     8581 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     8641 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     8701 gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg
     8761 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     8821 cctctagcta gagcttggcg taatcatggt catagatgtt tcctgtgtga aattgttatc
     8881 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     8941 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     9001 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     9061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     9121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     9181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     9241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     9301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     9361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     9421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     9481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     9541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     9601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     9661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     9721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     9781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     9841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     9901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     9961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    10021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    10081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    10141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    10201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    10261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    10321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    10381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    10441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    10501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    10561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    10621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    10681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    10741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    10801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    10861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    10921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    10981 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag
    11041 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
    11101 c
//
