LOCUS       sequence-447641-        9033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for ABE8e using SpCas9 enzyme 
            with MRKCRS amino acids at positions 1135,1136,1218,1219,1335,T1337 
            with C-term BPNLS-P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9033)
  AUTHORS   Silverstein RA, Kim N, Kroell AS, Walton RT, Delano J, Butcher RM, 
            Pacesa M, Smith BK, Christie KA, Ha LL, Meis RJ, Clark AB, Spinner 
            AD, Lazzarotto CR, Li Y, Matsubara A, Urbina EO, Dahl GA, Correia 
            BE, Marks DS, Tsai SQ, Pinello L, De Ravin SS, Liu Q, Kleinstiver BP
  TITLE     Custom CRISPR-Cas9 PAM variants via scalable engineering and machine
            learning.
  JOURNAL   Nature. 2025 Apr 22. doi: 10.1038/s41586-025-09021-y.
  PUBMED    40262634
REFERENCE   2  (bases 1 to 9033)
  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: "Nature. 
            2025 Apr 22. doi: 10.1038/s41586-025-09021-y."
COMMENT     Sequence Label: pCMV-T7-ABE8e-SpCas9(MRKCRS)-P2A-EGFP (BKS1034)
FEATURES             Location/Qualifiers
     source          1..9033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        160..539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        540..743
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     694..714
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     785..804
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        785..803
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             853..873
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1468..5568
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFMRPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKCLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYSSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5611..5631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5641..5697
                     /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(5698..5700,5701..5703,5704..6414)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5698..5700,5701..5703,5704..6414)
                     /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(6004..6023)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6351..6372
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6427..6447
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6456..6473
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6496..6513)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6502..6726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6797..6813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6797..6813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6810..6832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6821..6837
                     /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(6845..6851,6852..6869,6870..6875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6890..6911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7028..7045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7199..7787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7279..7298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7958..8749,8750..8818))
                     /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     8581..8600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8819..8923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9002..9021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
       61 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      121 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      181 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      241 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      301 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      361 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      421 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      481 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      541 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      601 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      661 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      721 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag agatccgcgg
      781 ccgctaatac gactcactat agggagagcc gccaccatga aacggacagc cgacggaagc
      841 gagttcgagt caccaaagaa gaagcggaaa gtctctgagg tggagttttc ccacgagtac
      901 tggatgagac atgccctgac cctggccaag agggcacgcg atgagaggga ggtgcctgtg
      961 ggagccgtgc tggtgctgaa caatagagtg atcggcgagg gctggaacag agccatcggc
     1021 ctgcacgacc caacagccca tgccgaaatt atggccctga gacagggcgg cctggtcatg
     1081 cagaactaca gactgattga cgccaccctg tacgtgacat tcgagccttg cgtgatgtgc
     1141 gccggcgcca tgatccactc taggatcggc cgcgtggtgt ttggcgtgag gaacagcaaa
     1201 agaggcgccg caggctccct gatgaacgtg ctgaactacc ccggcatgaa tcaccgcgtc
     1261 gaaattaccg agggaatcct ggcagatgaa tgtgccgccc tgctgtgcga tttctaccgg
     1321 atgcctagac aggtgttcaa tgctcagaag aaggcccaga gctccatcaa ctccggagga
     1381 tctagcggag gctcctctgg ctctgagaca cctggcacaa gcgagagcgc aacacctgaa
     1441 agcagcgggg gcagcagcgg ggggtcagac aagaagtaca gcatcggcct ggccatcggc
     1501 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     1561 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     1621 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     1681 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     1741 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     1801 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     1861 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     1921 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     1981 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     2041 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     2101 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     2161 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     2221 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     2281 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     2341 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     2401 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     2461 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     2521 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     2581 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     2641 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     2701 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     2761 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     2821 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     2881 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     2941 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     3001 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     3061 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     3121 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     3181 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     3241 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     3301 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     3361 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     3421 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     3481 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     3541 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     3601 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     3661 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     3721 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     3781 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     3841 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     3901 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     3961 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     4021 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     4081 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     4141 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     4201 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     4261 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     4321 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     4381 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     4441 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     4501 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     4561 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     4621 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     4681 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     4741 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     4801 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     4861 ggcttcatga gacccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     4921 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     4981 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     5041 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     5101 atgctggcct ctgccaagtg cctgcagaag ggaaacgaac tggccctgcc ctccaaatat
     5161 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     5221 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     5281 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     5341 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     5401 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     5461 cggaagagat acagcagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     5521 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggtgactc tggcggctca
     5581 aaaagaaccg ccgacggcag cgaattcgag cccaagaaga agaggaaagt cggaagcgga
     5641 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg
     5701 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5761 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     5821 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     5881 gtgaccaccc tgacctatgg agtgcagtgc ttcagccgct accccgacca catgaagcag
     5941 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     6001 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     6061 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     6121 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     6181 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     6241 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     6301 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     6361 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtctggt
     6421 ggttctccca agaagaagag gaaagtctaa ccggtcatca tcaccatcac cattgagttt
     6481 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     6541 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     6601 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     6661 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     6721 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
     6781 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6841 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
     6901 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6961 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7021 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7081 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7141 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7201 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7261 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7321 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7381 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7441 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7501 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7561 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7621 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7681 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7741 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7801 tcttttctac ggggtctgac actcagtgga acgaaaactc acgttaaggg attttggtca
     7861 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7921 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7981 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8041 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8101 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8161 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8221 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8281 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8341 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8401 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8461 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8521 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8581 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8641 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8701 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8761 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8821 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8881 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8941 tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg tcgactctca
     9001 gtacaatctg ctctgatgcc gcatagttaa gcc
//
