LOCUS       sequence-450493-        9744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses evoBE4max-SpRY base editor in mammalian cells with eGFP 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9744)
  AUTHORS   Osgood NRB, Zawalick NM, Sawyer CB, Cowan QT, Gu S, Mawson SJ, 
            Ranzau BL, Li L, Gymrek M, Goren A, Komor AC
  TITLE     Genome editing with programmable base editors in human cells
  JOURNAL   Methods in Enzymology, February 5, 2025
REFERENCE   2  (bases 1 to 9744)
  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: "Methods in 
            Enzymology, February 5, 2025"
COMMENT     Sequence Label: evoBE4-SpRY
FEATURES             Location/Qualifiers
     source          1..9744
                     /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     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             694..714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1495..5595
                     /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
                     LIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTRLGAPRAFKYFDTTIDPKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5626..5874
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5905..6153
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6196..6216
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6226..6282
                     /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(6283..6285,6286..6288,6289..7002)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6283..6285,6286..6288,6289..6999)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6589..6608)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6936..6957
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7008..7025
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7048..7065)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7054..7278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7349..7365)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7349..7365)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7362..7384)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7373..7389
                     /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(7397..7403,7404..7421,7422..7427))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7442..7463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7580..7597)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7751..8339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7831..7850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8510..9301,9302..9370))
                     /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     9133..9152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9371..9475)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9554..9573)
                     /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 agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcagttca
      721 aagactgggc ctgtcgccgt cgatccaacc ctgcgccgcc ggattgaacc tcacgagttt
      781 gaagtgttct ttgacccccg ggagctgaga aaggagacat gcctgctgta cgagatcaac
      841 tggggaggca ggcactccat ctggaggcac acctctcaga acacaaataa gcacgtggag
      901 gtgaacttca tcgagaagtt taccacagag cggtacttct gccccaatac cagatgtagc
      961 atcacatggt ttctgagctg gtccccttgc ggagagtgta gcagggccat caccgagttc
     1021 ctgtccagat atccaaatgt gacactgttt atctacatcg ccaggctgta tcacctggca
     1081 aacccaagga ataggcaggg cctgcgcgat ctgatcagct ccggcgtgac catccagatc
     1141 atgacagagc aggagtccgg ctactgctgg cacaacttcg tgaattattc tcctagcaac
     1201 gagtcccact ggcctaggta cccacacctg tgggtgcgcc tgtacgtgct ggagctgtat
     1261 tgcatcatcc tgggcctgcc cccttgtctg aatatcctgc ggagaaagca gagccagctg
     1321 acctccttta caatcgccct gcagtcttgt cactatcaga ggctgccacc ccacatcctg
     1381 tgggccacag gcctgaagtc tggcggatct agcggaggat cctctggcag cgagacacca
     1441 ggaacaagcg agtcagcaac accagagagc agtggcggca gcagcggcgg cagcgacaag
     1501 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
     1561 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     1621 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga gagaacccgg
     1681 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     1741 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     1801 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     1861 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     1921 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     1981 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     2041 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     2101 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     2161 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     2221 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     2281 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     2341 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     2401 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     2461 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     2521 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     2581 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     2641 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     2701 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     2761 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     2821 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     2881 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     2941 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     3001 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     3061 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     3121 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     3181 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     3241 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     3301 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     3361 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     3421 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     3481 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     3541 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     3601 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     3661 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     3721 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     3781 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     3841 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     3901 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     3961 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     4021 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     4081 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     4141 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     4201 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     4261 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     4321 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     4381 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     4441 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     4501 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     4561 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     4621 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     4681 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     4741 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     4801 ggcttcagca aagagtctat cagacccaag aggaacagcg ataagctgat cgccagaaag
     4861 aaggactggg accctaagaa gtacggcggc ttcctgtggc ccaccgtggc ctattctgtg
     4921 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     4981 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     5041 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     5101 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccaagcagct gcagaaggga
     5161 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     5221 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     5281 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     5341 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     5401 gagcaggccg agaatatcat ccacctgttt accctgacca gactgggagc ccctagagcc
     5461 ttcaagtact ttgacaccac catcgacccc aagcagtaca gaagcaccaa agaggtgctg
     5521 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     5581 cagctgggag gtgacagcgg cgggagcggc gggagcgggg ggagcactaa tctgagcgac
     5641 atcattgaga aggagactgg gaaacagctg gtcattcagg agtccatcct gatgctgcct
     5701 gaggaggtgg aggaagtgat cggcaacaag ccagagtctg acatcctggt gcacaccgcc
     5761 tacgacgagt ccacagatga gaatgtgatg ctgctgacct ctgacgcccc cgagtataag
     5821 ccttgggccc tggtcatcca ggattctaac ggcgagaata agatcaagat gctgagcgga
     5881 ggatccggag gatctggagg cagcaccaac ctgtctgaca tcatcgagaa ggagacaggc
     5941 aagcagctgg tcatccagga gagcatcctg atgctgcccg aagaagtcga agaagtgatc
     6001 ggaaacaagc ctgagagcga tatcctggtc cataccgcct acgacgagag taccgacgaa
     6061 aatgtgatgc tgctgacatc cgacgcccca gagtataagc cctgggctct ggtcatccag
     6121 gattccaacg gagagaacaa aatcaaaatg ctgtctggcg gctcaaaaag aaccgccgac
     6181 ggcagcgaat tcgagcccaa gaagaagaga aaagtcggaa gcggagctac taacttcagc
     6241 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatggtgag caagggcgag
     6301 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     6361 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     6421 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     6481 tatggagtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     6541 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     6601 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     6661 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     6721 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     6781 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     6841 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     6901 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     6961 gccgccggga tcactctcgg catggacgag ctgtacaagt aaccggtcat catcaccatc
     7021 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7081 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7141 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7201 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7261 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     7321 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7381 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     7441 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7501 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7561 ttgggcgcac ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7621 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca gaggataacg
     7681 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7741 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7801 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7861 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7921 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7981 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8041 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8101 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8161 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8221 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8281 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8341 aagatccttt gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag
     8401 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8461 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8521 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8581 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8641 tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8701 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8761 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8821 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8881 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8941 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9001 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9061 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9121 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9181 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9241 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9301 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9361 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9421 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9481 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag
     9541 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     9601 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     9661 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     9721 atgtacgggc cagatatacg cgtt
//
