LOCUS       sequence-225243-       10220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor construct to amplify 
            HA-NLS-dCas9-NLS-SID4x-TaV-2a-Citrine-FRT-Kan-FRT cassette for BAC 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10220)
  AUTHORS   Chong-Morrison V, Mayes S, Simoes FC, Senanayake U, Carroll DS, 
            Riley PR, Wilson SW, Sauka-Spengler T
  TITLE     Ac/Ds transposition for CRISPR/dCas9-SID4x epigenome modulation in 
            zebrafish.
  JOURNAL   Biol Open. 2023 Jun 15;12(6):bio059995. doi: 10.1242/bio.059995. 
            Epub 2023 Jun 27.
  PUBMED    37367831
REFERENCE   2  (bases 1 to 10220)
  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: "Biol 
            Open."; date: "2023-06-15"; volume: "12(6):bio059995. doi"; pages: "
            10.1242/bio.059995. Epub 2023 Jun 27"
COMMENT     Sequence Label: 
            pGEM-T-Easy-HA-NLS-dCas9-NLS-SID4x-TaV-2a-Citrine-FRT-Kan-FRT
FEATURES             Location/Qualifiers
     source          1..10220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      50..59
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      54..63
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             72..98
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             102..122
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(132..155,156..158,159..2714,2715..2717,2718..4232)
                     /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(132..155,156..158,159..2645,2646..2648,2649..4232)
                     /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             4236..4256
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4677..4730
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4737..4739,4740..4742,4743..5453)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5043..5062)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5390..5411
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5508..5629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5519..5538)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5573..5592
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(5753..5800)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        5894..5998
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6000..6190
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6023..6043)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             6230..7024
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6230..7024
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6284..6303)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6894..6913
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7031..7079
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    complement(7104..7151)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(7362..7380)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7363..7380)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7398..7414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7398..7414)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7411..7433)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7422..7438
                     /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(7446..7452,7453..7470,7471..7476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7491..7512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7629..7646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7800..8388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7880..7899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8559..9350,9351..9419))
                     /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     9182..9201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9420..9524)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9602..10057)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9739..9760)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9951..9970
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10183..10205
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10197..10214
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10198..10214
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc
       61 ggccgcggat gtacccatac gatgttccag attacgcttc gccgaagaaa aagcgcaagg
      121 tcgaagcgtc cgacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct
      181 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
      241 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
      301 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
      361 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
      421 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
      481 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
      541 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
      601 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
      661 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
      721 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
      781 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
      841 tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
      901 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
      961 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     1021 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     1081 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     1141 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     1201 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     1261 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     1321 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     1381 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     1441 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     1501 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     1561 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     1621 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     1681 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     1741 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     1801 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     1861 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     1921 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     1981 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     2041 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     2101 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     2161 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     2221 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     2281 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     2341 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     2401 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     2461 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     2521 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     2581 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     2641 tggacgccat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     2701 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     2761 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     2821 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     2881 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     2941 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     3001 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     3061 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     3121 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     3181 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     3241 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     3301 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     3361 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     3421 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     3481 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     3541 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     3601 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     3661 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     3721 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     3781 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     3841 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     3901 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     3961 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     4021 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     4081 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     4141 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     4201 cacggatcga cctgtctcag ctgggaggcg acagccccaa gaagaagaga aaggtggagg
     4261 ccagcgggcc ggccgaggcc agcatgaaca tccagatgct gctggaggcc gctgactacc
     4321 tggaacggag agagcgcgaa gccgagcacg gatatgcttc aatgctgccc ggaagcggca
     4381 tgaatattca gatgctgctg gaggctgctg attacctgga aaggcgcgaa cgggaggccg
     4441 aacatggcta tgcttccatg ctgcctgggt ctggaatgaa tatccaaatg ctgctggagg
     4501 cagccgatta cctggaacgg agagaaagag aagccgagca cggatacgcc agcatgctgc
     4561 caggcagcgg gatgaacata caaatgctgc tggaggctgc cgattacctg gagaggcgcg
     4621 agagagaagc tgaacatggc tatgcctcta tgctgcccct cgagatcgga tcaggagagg
     4681 gcaggggaag tcttctaaca tgcggggacg tggaggaaaa tcccgggccc agatctatgg
     4741 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     4801 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     4861 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     4921 tgaccacctt cggctacggc ctgatgtgct tcgcccgcta ccccgaccac atgaagcagc
     4981 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     5041 aagacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     5101 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     5161 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     5221 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     5281 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     5341 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     5401 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     5461 gccgcgactc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     5521 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5581 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5641 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     5701 aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgacctgc ttgaagttcc
     5761 tatactttct agagaatagg aacttcggaa taggaacttc aagatccccc tggcgaaagg
     5821 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcgtcag gtggcacttt
     5881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     6001 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     6061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     6121 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagta
     6181 accatagtcc gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6241 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6301 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6361 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6421 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6481 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6541 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6601 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6661 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6721 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     6781 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6841 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6901 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6961 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     7021 ctgaggggat cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttcg
     7081 gatccgagct tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga
     7141 ataggaactt ctccatggta gcctccaaaa aagcctcctc actacttctg gaaaagctca
     7201 atcagtaagt acaaaataat gataatgaaa aaaagagcta tatgtatatg atagttatac
     7261 atatgtaggc ccgcacggaa taatcactag tgaattcgcg gccgcctgca ggtcgaccat
     7321 atgggagagc tcccaacgcg ttggatgcat agcttgagta ttctatagtg tcacctaaat
     7381 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7441 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7501 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7561 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     9001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     9061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     9121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     9181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9541 gtgccacctg atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     9601 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     9661 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     9721 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     9781 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     9841 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     9901 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     9961 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
    10021 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
    10081 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
    10141 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
    10201 aaacgacggc cagtgaattg
//
