LOCUS       sequence-234985-       12598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter driven expression of ABEmax base editor, EGFP and 
            Puromycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12598)
  TITLE     Otonkoski plasmids 2018
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12598)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CAG-ABEmax-P2A-EGFP-ires-puro
FEATURES             Location/Qualifiers
     source          1..12598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    304..438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        452..831
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        833..1110
                     /label=chicken beta-actin promoter
     intron          1111..2127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2051..2073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2151..2170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2151..2169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2219..2239
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3428..3451,3452..3454,3455..7528)
                     /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             3428..7528
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7571..7591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7601..7657
                     /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(7658..7660,7661..7663,7664..8377)
                     /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(7658..7660,7661..7663,7664..8374)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7964..7983)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8311..8332
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(8399..8973,8974..8976,8977..8985)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8566..8583)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8793..8812
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8986..9585
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8986..9005)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9482..9502
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(9666..9683)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9672..9896
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9943..9959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9943..9959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9956..9978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9967..9983
                     /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(9991..9997,9998..10015,10016..10021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10036..10057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10116..10311
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      10162..10297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    10317..10451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10519..10536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10690..11278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10770..10789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11449..12240,12241..12309))
                     /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     12072..12091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgtcatagt aatgattggg tggaaacatt ccaggcctgg gtggagaggc tttttgcttc
       61 ctcttgcaaa accacactga cattccaggc ctgggtggag aggctttttg cttcctcttg
      121 caaaaccaca ctgccctctg gagggcagtt gcctagcaac taattaaaag aggatgtcgc
      181 acggccagct gcggtcagtt agtcacttcc tgcttaactg acttgacatt ttctatttta
      241 agagtcggga ggaaaattac tgtgttggag gccctccgcc atcttctgaa gctgaatcga
      301 attaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
      361 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
      421 tcttatcatg tctggatctg atatcatcgt cgacattgat tattgactag ttattaatag
      481 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      541 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      601 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      661 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      721 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      781 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
      841 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
      901 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
      961 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     1021 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     1081 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
     1141 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     1201 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1261 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
     1321 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
     1381 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1441 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc
     1501 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc
     1561 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg
     1621 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg
     1681 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg
     1741 gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat
     1801 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
     1861 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
     1921 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
     1981 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     2041 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     2101 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggccgc taatacgact
     2161 cactataggg agagccgcca ccatgaaacg gacagccgac ggaagcgagt tcgagtcacc
     2221 aaagaagaag cggaaagtct ctgaagtcga gtttagccac gagtattgga tgaggcacgc
     2281 actgaccctg gcaaagcgag catgggatga aagagaagtc cccgtgggcg ccgtgctggt
     2341 gcacaacaat agagtgatcg gagagggatg gaacaggcca atcggccgcc acgaccctac
     2401 cgcacacgca gagatcatgg cactgaggca gggaggcctg gtcatgcaga attaccgcct
     2461 gatcgatgcc accctgtatg tgacactgga gccatgcgtg atgtgcgcag gagcaatgat
     2521 ccacagcagg atcggaagag tggtgttcgg agcacgggac gccaagaccg gcgcagcagg
     2581 ctccctgatg gatgtgctgc accaccccgg catgaaccac cgggtggaga tcacagaggg
     2641 aatcctggca gacgagtgcg ccgccctgct gagcgatttc tttagaatgc ggagacagga
     2701 gatcaaggcc cagaagaagg cacagagctc caccgactct ggaggatcta gcggaggatc
     2761 ctctggaagc gagacaccag gcacaagcga gtccgccaca ccagagagct ccggcggctc
     2821 ctccggagga tcctctgagg tggagttttc ccacgagtac tggatgagac atgccctgac
     2881 cctggccaag agggcacgcg atgagaggga ggtgcctgtg ggagccgtgc tggtgctgaa
     2941 caatagagtg atcggcgagg gctggaacag agccatcggc ctgcacgacc caacagccca
     3001 tgccgaaatt atggccctga gacagggcgg cctggtcatg cagaactaca gactgattga
     3061 cgccaccctg tacgtgacat tcgagccttg cgtgatgtgc gccggcgcca tgatccactc
     3121 taggatcggc cgcgtggtgt ttggcgtgag gaacgcaaaa accggcgccg caggctccct
     3181 gatggacgtg ctgcactacc ccggcatgaa tcaccgcgtc gaaattaccg agggaatcct
     3241 ggcagatgaa tgtgccgccc tgctgtgcta tttctttcgg atgcctagac aggtgttcaa
     3301 tgctcagaag aaggcccaga gctccaccga ctccggagga tctagcggag gctcctctgg
     3361 ctctgagaca cctggcacaa gcgagagcgc aacacctgaa agcagcgggg gcagcagcgg
     3421 ggggtcagac aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc
     3481 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     3541 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     3601 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     3661 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     3721 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     3781 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     3841 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     3901 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     3961 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     4021 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     4081 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     4141 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     4201 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     4261 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     4321 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     4381 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     4441 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     4501 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     4561 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     4621 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     4681 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     4741 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     4801 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     4861 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     4921 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     4981 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     5041 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     5101 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     5161 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     5221 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     5281 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     5341 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     5401 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     5461 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     5521 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     5581 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     5641 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     5701 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     5761 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     5821 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     5881 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     5941 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     6001 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     6061 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     6121 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     6181 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     6241 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     6301 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     6361 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     6421 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     6481 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     6541 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     6601 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     6661 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     6721 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     6781 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     6841 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     6901 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     6961 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     7021 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     7081 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     7141 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     7201 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     7261 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     7321 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     7381 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     7441 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     7501 gatcgacctg tctcagctgg gaggtgactc tggcggctca aaaagaaccg ccgacggcag
     7561 cgaattcgag cccaagaaga agaggaaagt cggaagcgga gctactaact tcagcctgct
     7621 gaagcaggct ggagacgtgg aggagaaccc tggacctatg gtgagcaagg gcgaggagct
     7681 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     7741 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     7801 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctatgg
     7861 agtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     7921 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     7981 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     8041 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     8101 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     8161 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     8221 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     8281 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     8341 cgggatcact ctcggcatgg acgagctgta caagtaattt aaagaattcc gccccccccc
     8401 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     8461 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     8521 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     8581 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     8641 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     8701 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     8761 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     8821 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     8881 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     8941 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatgac cgagtacaag
     9001 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     9061 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     9121 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     9181 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     9241 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     9301 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     9361 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     9421 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     9481 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     9541 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     9601 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgatgatcta gagctcgctg
     9661 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     9721 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     9781 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     9841 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     9901 tgaggcggaa agaacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     9961 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    10021 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    10081 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
    10141 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
    10201 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
    10261 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
    10321 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
    10381 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
    10441 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    10501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    10561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    10621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    10681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    10741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    10801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    10861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    10921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    10981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    11041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    11101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    11161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    11221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    11281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    11341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    11401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    11461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    11521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    11581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    11641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    11701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    11761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    11821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    11881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    11941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    12001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    12061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    12121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    12181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    12241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    12301 aatactcata ctcttccttt ttcaatgacc gctttggaaa aacaaagact gtatttcctg
    12361 gaaattaatg tttattcaat aaactgtgta ttcagctata ttcacatagt ggtgaggctg
    12421 aaatgaggcg ggaagaggcg gttggggctt aattatatca atttgggtgg ccccacagcg
    12481 cctccaaggc gccagtcctg ttttgacaag ttgcctctgg aagcctctct tctttttctc
    12541 cagagtaagc ggaggccagg ggcccccggc ctctgcttaa tactaaaaaa aacagctg
//
