LOCUS       sequence-454996-        9533 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Using promoter ef1a expresses ABE8e-dCas9 protein with Blasticidin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9533)
  TITLE     Plasmid2MC: Efficient cell-free recombination of plasmids into 
            high-purity minicircle DNA for use in genome editing applications
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9533)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: p-att-ef1a-ABE8e-dCas9-BSD
FEATURES             Location/Qualifiers
     source          1..9533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    43..76
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        185..1363
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          416..1354
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1311..1331
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1412..1432
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2027..2050,2051..2053,2054..4609,4610..4612,
                     4613..6127)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(2027..2050,2051..2053,2054..4540,4541..4543,
                     4544..6127)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6128..6175
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6185..6241
                     /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             6242..6640
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(6668..6685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6674..6881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7040..7495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7127..7146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7337..7358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7512..7531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7631..7653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7691..7709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7777..7881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7882..7950,7951..8742)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8100..8119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8913..9501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9402..9421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tggccttttg ctcacgagac tcagcggccg cggtgccagg gcgtgccctt
       61 gggctccccg ggcgcgacta gtgaattcag atctgatatc tctgtcggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      181 gtccggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
      241 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
      301 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
      361 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
      421 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
      481 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
      541 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
      601 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
      661 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
      721 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
      781 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
      841 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
      901 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
      961 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     1021 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     1081 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     1141 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     1201 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     1261 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     1321 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgaa
     1381 acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag tctctgaggt
     1441 ggagttttcc cacgagtact ggatgagaca tgccctgacc ctggccaaga gggcacggga
     1501 tgagagggag gtgcctgtgg gagccgtgct ggtgctgaac aatagagtga tcggcgaggg
     1561 ctggaacaga gccatcggcc tgcacgaccc aacagcccat gccgaaatta tggccctgag
     1621 acagggcggc ctggtcatgc agaactacag actgattgac gccaccctgt acgtgacatt
     1681 cgagccttgc gtgatgtgcg ccggcgccat gatccactct aggatcggcc gcgtggtgtt
     1741 tggatggaga aattctaaaa gaggcgccgc aggctccctg atgaacgtgc tgaactaccc
     1801 cggcatgaat caccgcgtcg aaattaccga gggaatcctg gcagatgaat gtgccgccct
     1861 gctgtgcgat ttctatcgga tgcctagaca ggtgttcaat gctcagaaga aggcccagag
     1921 ctccatcaac tccggaggat ctagcggagg ctcctctggc tctgagacac ctggcacaag
     1981 cgagagcgca acacctgaaa gcagcggggg cagcagcggg gggtcagaca agaagtacag
     2041 catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     2101 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     2161 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     2221 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     2281 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     2341 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     2401 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     2461 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     2521 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     2581 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     2641 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     2701 cgcccagctg cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct
     2761 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     2821 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     2881 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     2941 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     3001 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     3061 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     3121 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     3181 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     3241 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     3301 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     3361 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     3421 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     3481 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     3541 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     3601 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     3661 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     3721 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     3781 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     3841 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     3901 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     3961 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     4021 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     4081 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     4141 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     4201 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     4261 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     4321 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     4381 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     4441 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     4501 ggaactggac atcaaccggc tgtccgacta cgatgtggac gccatcgtgc ctcagagctt
     4561 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     4621 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     4681 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     4741 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     4801 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     4861 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     4921 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     4981 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     5041 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     5101 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     5161 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     5221 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     5281 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     5341 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     5401 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     5461 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     5521 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     5581 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     5641 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     5701 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     5761 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     5821 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     5881 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     5941 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     6001 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     6061 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     6121 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagggaag
     6181 cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc
     6241 tatggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     6301 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     6361 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     6421 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     6481 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     6541 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     6601 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gaattctaac tagagctcgc
     6661 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6721 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6781 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6841 aagggggagg attgggaaga gaatagcagg catgctgggg acctgcagga gctcgtcgac
     6901 ccatgggggc ccgccccaac tggggtaacc tttgagttct ctcagttggg ggtaatcagc
     6961 atcatgatgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     7021 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7081 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     7141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     7261 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     7381 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     7441 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     7501 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     7561 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     7621 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     7681 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     7741 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     7801 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7861 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7921 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     7981 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     8041 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     8101 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     8161 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     8221 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     8281 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     8341 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     8401 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     8461 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     8521 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8581 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     8641 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8701 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     8761 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8821 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     8941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     9001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     9061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     9121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     9181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     9241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     9301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     9361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     9421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     9481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cct
//
