LOCUS       sequence-215839-       10634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses D10A-GCN4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10634)
  AUTHORS   Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang 
            L, Hou Y, Chen J, Chen J, Huang X
  TITLE     BE-PLUS: a new base editing tool with broadened editing window and 
            enhanced fidelity.
  JOURNAL   Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 
            10.1038/s41422-018-0052-4.
  PUBMED    29875396
REFERENCE   2  (bases 1 to 10634)
  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: "Cell Res. 
            2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 
            10.1038/s41422-018-0052-4."
COMMENT     Sequence Label: pST1374-D10A-GCN4
FEATURES             Location/Qualifiers
     source          1..10634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             676..696
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             706..729
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(736..759,760..762,763..4836)
                     /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             4867..4887
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4894..4914
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4951..5007
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5089..5145
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5227..5283
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5365..5421
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5503..5559
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5641..5697
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5779..5835
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5917..5973
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6055..6111
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6193..6249
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6289..6309
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6316..6333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6356..6373)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6362..6586
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6632..7060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6719..6738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6929..6950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7069..7089)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7074..7403
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7254..7389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7316..7335
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7451..7498
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7517..7915
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8073..8194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8110..8129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8164..8183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8243..8259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8243..8259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8256..8278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8267..8283
                     /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(8291..8297,8298..8315,8316..8321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8336..8357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8474..8491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8645..9233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8725..8744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9404..10195,10196..10264))
                     /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     10027..10046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10265..10369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10444..10463)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcacca tgggacctaa gaaaaagagg aaggtggcgg ccgctgacta caaggatgac
      721 gacgataaat ctagagacaa gaaatactct attggactgg ctatcgggac aaactccgtt
      781 ggctgggccg tcataaccga cgagtataag gtgccaagca agaaattcaa ggtgctgggt
      841 aatactgacc gccattcaat caagaagaac ctgatcggag cactcctctt cgactccggt
      901 gaaaccgctg aagctactcg gctgaagcgg accgcaaggc ggagatacac ccgccgcaag
      961 aatcggatat gttatctgca agagatcttt agcaacgaaa tggctaaggt ggacgactcc
     1021 ttctttcacc gcctggaaga gagctttctg gtggaggagg ataagaaaca cgagaggcac
     1081 cctatattcg gaaatatcgt ggatgaggtg gcttaccatg aaaagtatcc tacaatctac
     1141 catctgagga agaagctggt ggacagcacc gataaagcag acctgaggct catctatctg
     1201 gccctggctc atatgataaa gtttagagga cactttctga tcgagggcga cctgaatccc
     1261 gataattccg atgtggataa actcttcatt caactggtgc agacatataa ccaactgttc
     1321 gaggagaatc ccataaacgc ttctggtgtg gatgccaagg ctattctgtc cgctcggctg
     1381 tccaagtcac gcagactgga gaatctgatt gcccaactgc caggagaaaa gaagaacggc
     1441 ctgtttggga acctcatcgc cctgagcctg ggcctgacac ctaacttcaa gtccaatttt
     1501 gatctggccg aagatgctaa actccagctc tccaaggaca cctatgacga tgatctggac
     1561 aacctgctcg cacagatagg cgaccagtac gccgatctct ttctggctgc taagaatctc
     1621 tccgacgcca ttctgctgag cgacatactc cgggtcaaca ctgagatcac caaagcacct
     1681 ctgagcgcct ccatgataaa acgctatgat gaacaccatc aagacctgac tctgctcaaa
     1741 gccctcgtga ggcaacagct gccagagaag tacaaagaga tattcttcga ccagagcaag
     1801 aatggatatg ccggatacat cgatggcgga gcatcacagg aagaatttta caagttcatc
     1861 aaaccaatcc tcgagaagat ggacggtact gaagagctgc tggtgaagct gaacagggag
     1921 gacctgctga ggaagcagag gacctttgat aatggctcca ttccacatca gatacacctg
     1981 ggagagctgc atgcaatcct ccgcaggcag gaggatttct atcctttcct gaaggataac
     2041 cgggagaaga tagagaagat cctgaccttc aggatccctt attacgtcgg ccctctggct
     2101 agaggcaact cccgcttcgc ttggatgacc aggaaatctg aggagacaat tactccttgg
     2161 aacttcgaag aggtcgtgga taagggcgca agcgcccagt cattcatcga acggatgacc
     2221 aatttcgata agaacctgcc caacgagaag gtcctgccca aacattcact cctgtacgag
     2281 tatttcaccg tctataacga gctgactaaa gtgaagtacg tgaccgaggg catgaggaag
     2341 cctgccttcc tgtccggaga gcagaagaag gctatcgttg atctgctctt caagactaat
     2401 agaaaggtga cagtgaagca gctcaaggag gattacttta agaagatcga atgctttgac
     2461 tcagtggaaa tctctggcgt ggaggaccgc tttaatgcca gcctgggcac ttaccatgat
     2521 ctgctgaaga taatcaaaga caaagatttc ctcgataatg aggagaacga ggacatcctg
     2581 gaagatatcg tgctgaccct gactctgttc gaggatagag agatgatcga agagcgcctg
     2641 aagacctatg cccatctgtt tgacgataaa gtcatgaaac agctcaagcg gcggcgctac
     2701 actgggtggg gtagactctc caggaaactc ataaacggca tccgcgacaa acagagcgga
     2761 aagaccatcc tggatttcct gaaatccgac ggattcgcta acaggaactt catgcaactg
     2821 attcacgatg actctctgac atttaaagag gacatccaga aggcacaggt gagcggtcaa
     2881 ggcgacagcc tgcacgagca catcgccaac ctcgctggat cacccgccat aaagaaggga
     2941 atactgcaga cagtcaaggt cgtggacgaa ctcgtcaaag tgatgggtcg gcacaagcca
     3001 gagaatatcg ttatcgaaat ggcaagggag aaccaaacca cccagaaggg ccagaagaac
     3061 tctcgggaac ggatgaaaag aatcgaagag ggaattaagg agctgggatc tcagatactg
     3121 aaggagcacc ctgtggagaa tacacagctc cagaacgaga aactctacct gtactacctc
     3181 cagaacgggc gggacatgta cgttgaccag gaactcgaca tcaaccggct gtccgattat
     3241 gacgtggacc atattgttcc acagtccttc ctcaaagatg actccattga caacaaggtg
     3301 ctgaccagat ccgataagaa tcgcggtaag tctgacaatg ttccatcaga agaggtggtc
     3361 aagaagatga agaattactg gcggcagctc ctcaacgcca aactgatcac ccagcggaag
     3421 tttgacaatc tgactaaggc agaaagagga ggtctgagcg aactcgacaa ggccggcttt
     3481 attaagaggc aactggtcga aacacgccag attaccaaac acgtggcaca aatcctcgac
     3541 tctaggatga acactaagta cgatgagaac gataagctga tcagggaagt gaaagtgata
     3601 actctgaaga gcaagctggt gtctgacttc cggaaggact ttcaattcta caaagttcgc
     3661 gaaataaaca attaccatca tgctcacgat gcctatctca atgctgtcgt tggcaccgcc
     3721 ctgatcaaga aataccctaa actggagtct gagttcgtgt acggtgacta taaagtctac
     3781 gatgtgagga agatgatagc aaagtctgag caagagattg gcaaagccac cgccaagtac
     3841 ttcttctact ctaatatcat gaatttcttt aagactgaga taaccctggc taacggcgaa
     3901 atccggaagc gcccactgat cgaaacaaac ggagaaacag gagaaatcgt gtgggataaa
     3961 ggcagggact tcgcaactgt gcggaaggtg ctgtccatgc cacaagtcaa tatcgtgaag
     4021 aagaccgaag tgcagaccgg cggattctca aaggagagca tcctgccaaa gcggaactct
     4081 gacaagctga tcgccaggaa gaaagattgg gacccaaaga agtatggcgg tttcgattcc
     4141 cctacagtgg cttattccgt tctggtcgtg gcaaaagtgg agaaaggcaa gtccaagaaa
     4201 ctcaagtctg ttaaggagct gctcggaatt actattatgg agagatccag cttcgagaag
     4261 aatccaatcg atttcctgga agctaagggc tataaagaag tgaagaaaga tctcatcatc
     4321 aaactgccca agtactctct ctttgagctg gagaatggta ggaagcggat gctggcctcc
     4381 gccggagagc tgcagaaagg aaacgagctg gctctgccct ccaaatacgt gaacttcctg
     4441 tatctggcct cccactacga gaaactcaaa ggtagccctg aagacaatga gcagaagcaa
     4501 ctctttgttg agcaacataa acactacctg gacgaaatca ttgaacagat tagcgagttc
     4561 agcaagcggg ttattctggc cgatgcaaac ctcgataaag tgctgagcgc atataataag
     4621 cacagggaca agccaattcg cgaacaagca gagaatatta tccacctctt tactctgact
     4681 aatctgggcg ctcctgctgc cttcaagtat ttcgatacaa ctattgacag gaagcggtac
     4741 acctctacca aagaagttct cgatgccacc ctgatacacc agtcaattac cggactgtac
     4801 gagactcgca tcgacctgtc tcagctcggc ggcgacggtt ctgaattcag cctgggcagc
     4861 ggctccccca agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa agtggacggc
     4921 attggtagtg ggagcaacgg cagcagcgga gaggaactgc tgagcaagaa ctaccacctg
     4981 gaaaacgagg tggccagact gaagaaaggc tctggctctg gcggaagcgg ttctggcgga
     5041 tcaggatctg gaagtggcgg ctctggaagc ggaggttctg gatctggcga agaactgctg
     5101 tctaagaatt atcacctcga gaacgaagtg gctcggctca agaaaggcag tggcagcgga
     5161 ggaagtggat ccggcggtag tggtagtgga agcggcggat ccggctcagg cggatctggt
     5221 tcaggggaag aactcctctc caaaaactac catctcgaga atgaggtcgc ccgcctgaaa
     5281 aaaggatcag gttccggtgg ttctggcagc ggtggttcag gctcaggcag cggcggtagc
     5341 ggtagcggtg gaagcggaag cggagaagaa cttctcagca aaaattacca cttggagaat
     5401 gaagttgcaa gactcaaaaa aggttccggc agtggcggca gcggcagcgg cggatctggt
     5461 agtggatctg gtggcagtgg ttcaggcgga agtggtagcg gagaggaatt gctctcaaag
     5521 aactatcatt tggagaacga ggttgcacgc ttgaagaaag gcagcggatc aggcggatct
     5581 ggcagcggtg gatctggttc tggatccggc ggctccggta gtggtggaag tggctctggg
     5641 gaagaattgc ttagcaagaa ttatcatctt gaaaatgagg ttgccaggct taaaaaaggc
     5701 agtggctccg gcggatccgg aagcggaggc agcggatctg gatctggtgg ttcaggatct
     5761 ggcggttctg gtagcgggga agaactgttg agtaaaaact atcaccttga gaacgaggtc
     5821 gcaaggttga aaaaaggatc cggctctggc ggctccggaa gtggcggatc tggctccggt
     5881 agcggaggat caggatccgg cggaagcgga tcaggcgagg aactgctttc caaaaattac
     5941 caccttgaaa acgaagtcgc ccgcctcaag aaaggttctg gcagcggagg ctctggcagt
     6001 ggtggtagcg gaagtggaag tggtggcagt ggtagcggtg gatctggaag cggcgaggaa
     6061 ctcctgtcaa agaattacca tctcgaaaac gaggtcgcaa ggctcaagaa aggctcagga
     6121 tcaggcggct ctggatccgg cggttctggt tccggctcag gtggaagtgg atctggcggc
     6181 tcaggttccg gcgaagaatt gctttccaag aactaccatt tggaaaatga agtcgctcgt
     6241 ttgaagaaag gttcaggctc cggccagcgg ccgcaaggtg gaggtggacc caagaagaag
     6301 cgcaaggtga ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg
     6361 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6421 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6481 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6541 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     6601 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     6661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6841 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6961 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     7081 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7141 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     7201 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     7261 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     7321 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     7381 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     7441 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     7501 ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     7561 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     7621 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     7681 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     7741 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     7801 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     7861 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagcact
     7921 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     7981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     8041 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     8101 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     8161 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     8221 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     8281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     8341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8401 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     9001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     9061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     9121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     9181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     9241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     9481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    10021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    10081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    10141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    10201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    10261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    10321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10381 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
    10441 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
    10501 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
    10561 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
    10621 cagatatacg cgtt
//
