LOCUS       sequence-235381-       12494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter driven expression of xFNLS BE3 base editor, EGFP and 
            Puromycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12494)
  TITLE     Otonkoski plasmids 2018
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12494)
  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            "Unpublished"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-xFNLS-T2A-EGFP-ires-puro
FEATURES             Location/Qualifiers
     source          1..12494
                     /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     2135..2154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2226..2235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2235..2300
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2307..2327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2352..3038
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             3087..7187
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 mutant with broad PAM
                     compatibility (Hu et al., 2018)"
                     /label=xCas9-3.7
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3087..7187
                     /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
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7200..7448
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7461..7481
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7497..7553
                     /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(7554..7556,7557..7559,7560..8273)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7860..7879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8207..8228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(8295..8869,8870..8872,8873..8881)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8462..8479)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8689..8708
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8882..9481
                     /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(8882..8901)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9378..9398
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(9562..9579)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9568..9792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9839..9855)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9839..9855)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9852..9874)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9863..9879
                     /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(9887..9893,9894..9911,9912..9917))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9932..9953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10012..10207
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      10058..10193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    10213..10347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10415..10432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10586..11174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10666..10685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11345..12136,12137..12205))
                     /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     11968..11987
                     /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 tgctggttat tgtgctgtct
     2161 catcattttg gcaaagaatt cctcgaccgg tgatcgcatg tcgacgctag cctctcagta
     2221 gatccgccac catggactat aaggaccacg acggagacta caaggatcat gatattgatt
     2281 acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag
     2341 tcccagcagc catgagctca gagactggcc cagtggctgt ggaccccaca ttgagacggc
     2401 ggatcgagcc ccatgagttt gaggtattct tcgatccgag agagctccgc aaggagacct
     2461 gcctgcttta cgaaattaat tgggggggcc ggcactccat ttggcgacat acatcacaga
     2521 acactaacaa gcacgtcgaa gtcaacttca tcgagaagtt cacgacagaa agatatttct
     2581 gtccgaacac aaggtgcagc attacctggt ttctcagctg gagcccatgc ggcgaatgta
     2641 gtagggccat cactgaattc ctgtcaaggt atccccacgt cactctgttt atttacatcg
     2701 caaggctgta ccaccacgct gacccccgca atcgacaagg cctgcgggat ttgatctctt
     2761 caggtgtgac tatccaaatt atgactgagc aggagtcagg atactgctgg agaaactttg
     2821 tgaattatag cccgagtaat gaagcccact ggcctaggta tccccatctg tgggtacgac
     2881 tgtacgttct tgaactgtac tgcatcatac tgggcctgcc tccttgtctc aacattctga
     2941 gaaggaagca gccacagctg acattcttta ccatcgctct tcagtcttgt cattaccagc
     3001 gactgccccc acacattctc tgggccaccg ggttgaaaag cggcagcgag actcccggga
     3061 cctcagagtc cgccacaccc gaaagtgaca agaagtacag catcggcctg gccatcggca
     3121 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     3181 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     3241 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     3301 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     3361 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     3421 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     3481 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     3541 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     3601 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     3661 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     3721 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     3781 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     3841 agagcaactt cgacctggcc gaggatacca aactgcagct gagcaaggac acctacgacg
     3901 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     3961 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     4021 ccaaggcccc cctgagcgcc tctatgatca agctgtacga cgagcaccac caggacctga
     4081 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     4141 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     4201 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     4261 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcatc atcccccacc
     4321 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     4381 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     4441 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     4501 tcaccccctg gaacttcgag aaggtggtgg acaagggcgc ttccgcccag agcttcatcg
     4561 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     4621 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     4681 gaatgagaaa gcccgccttc ctgagcggcg accagaaaaa ggccatcgtg gacctgctgt
     4741 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     4801 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     4861 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     4921 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     4981 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     5041 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     5101 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     5161 tcatccagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     5221 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     5281 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     5341 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     5401 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     5461 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     5521 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     5581 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
     5641 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     5701 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     5761 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     5821 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     5881 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     5941 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     6001 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     6061 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     6121 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     6181 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     6241 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     6301 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     6361 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     6421 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     6481 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     6541 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     6601 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     6661 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     6721 tgctggcctc tgccggcgtg ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     6781 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     6841 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     6901 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     6961 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     7021 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     7081 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     7141 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgattca ggcggatcta
     7201 ctaatctgtc agatattatt gaaaaggaga ccggtaagca actggttatc caggaatcca
     7261 tcctcatgct cccagaggag gtggaagaag tcattgggaa caagccggaa agcgatatac
     7321 tcgtgcacac cgcctacgac gagagcaccg acgagaatgt catgcttctg actagcgacg
     7381 cccctgaata caagccttgg gctctggtca tacaggatag caacggtgag aacaagatta
     7441 agatgctctc tggtggttct cccaagaaga agaggaaagt catgcatggc agcggcgcca
     7501 ccaacttcag cctgctgaag caggccggcg acgtggagga gaaccccggc cccatggtga
     7561 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     7621 taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc tacggcaagc
     7681 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     7741 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     7801 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     7861 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     7921 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     7981 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     8041 aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     8101 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     8161 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     8221 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taatttaaag
     8281 aattccgccc cccccccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
     8341 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     8401 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     8461 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     8521 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     8581 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     8641 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     8701 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     8761 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     8821 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     8881 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     8941 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     9001 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     9061 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     9121 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     9181 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     9241 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     9301 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     9361 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     9421 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     9481 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     9541 gatctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     9601 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     9661 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     9721 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     9781 tgggctctat ggcttctgag gcggaaagaa cctgcagccc aagcttggcg taatcatggt
     9841 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     9901 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     9961 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
    10021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
    10081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
    10141 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
    10201 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    10261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    10321 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
    10381 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
    10441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
    10501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
    10561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    10621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    10681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    10741 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    10801 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    10861 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    10921 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    10981 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
    11041 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    11101 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    11161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    11221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
    11281 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
    11341 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
    11401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
    11461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
    11521 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    11581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    11641 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    11701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    11761 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    11821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    11881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    11941 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    12001 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    12061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    12121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    12181 cgacacggaa atgttgaata ctcatactct tcctttttca atgaccgctt tggaaaaaca
    12241 aagactgtat ttcctggaaa ttaatgttta ttcaataaac tgtgtattca gctatattca
    12301 catagtggtg aggctgaaat gaggcgggaa gaggcggttg gggcttaatt atatcaattt
    12361 gggtggcccc acagcgcctc caaggcgcca gtcctgtttt gacaagttgc ctctggaagc
    12421 ctctcttctt tttctccaga gtaagcggag gccaggggcc cccggcctct gcttaatact
    12481 aaaaaaaaca gctg
//
