LOCUS       Ac5-STABLE2-neo-CAHS3        7791 bp    DNA     circular UNA 12-OCT-2021
DEFINITION  .
ACCESSION   urn.local...41-dyo0shf
VERSION     urn.local...41-dyo0shf
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    1..6
                     /Recognition_pattern="GGTAC^C"
                     /Methylation="Dcm:cut, CpG:cut"
                     /label="KpnI"
                     /note="Geneious type: restriction site"
     misc_feature    1..6
                     /Recognition_pattern="G^GTACC"
                     /Methylation="Mixed Methylation"
                     /label="Acc65I, Asp718I"
                     /note="Geneious type: restriction site"
     misc_feature    8..13
                     /Recognition_pattern="A^CTAGT"
                     /Methylation="Mixed Methylation"
                     /label="SpeI, AhlI, BcuI"
                     /note="Geneious type: restriction site"
     misc_feature    14..25
                     /Recognition_pattern="CCANNNNN^NTGG"
                     /Methylation="Dcm:variable"
                     /label="BstXI"
                     /note="Geneious type: restriction site"
     primer          complement(22..42)
                     /created_by="saibou"
                     /label="STABLE2-iF"
     misc_feature    25..30
                     /Recognition_pattern="G^AATTC"
                     /Methylation="CpG:variable, EcoBI:variable"
                     /label="EcoRI"
                     /note="Geneious type: restriction site"
     primer          28..64
                     /created_by="saibou"
                     /modified_by="saibou"
                     /label="C3-stable2-fw"
     regulatory      31..40
                     /regulatory_class="other"
                     /note="<html><body>vertebrate consensus sequence for
                     strong initiation of translation (<a
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/3313277""
                     title=""https://www.ncbi.nlm.nih.gov/pubmed/3313277"">Koza
                     k, 1987</a>)</body></html>"
                     /label="Kozak sequence"
     CDS             40..>42
                     /transl_table=1
                     /codon_start=1
                     /product="<html><body>FLAG® epitope tag, followed by an
                     enterokinase cleavage site</body></html>"
                     /Original_Translation="DYKDDDDK"
                     /label="FLAG"
     misc_feature    43..951
                     /Original_Bases="TACAAAGACGATGACGACAAGGGCAGCGTGTCCAAGGGCGA
                     AGAGGACAACATGGCCATCATCAAGGAGTTCATGCGGTTCAAGGTGCACATGGAAGGC
                     AGCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAAGGCAGGCCCTACGAGG
                     GCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGCGGACCTCTGCCCTTCGCCTGGGA
                     CATCCTGAGCCCCCAGTTCATGTACGGCAGCAAGGCCTACGTGAAGCACCCCGCCGAC
                     ATCCCCGACTACCTGAAGCTGTCCTTCCCCGAGGGATTCAAATGGGAGCGGGTGATGA
                     ACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACAGCAGCCTGCAGGACGGCGA
                     GTTCATCTACAAGGTGAAACTGCGGGGCACCAACTTCCCCAGCGACGGCCCCGTGATG
                     CAGAAAAAGACCATGGGCTGGGAGGCCAGCAGCGAGCGGATGTACCCCGAGGATGGCG
                     CCCTGAAGGGCGAGATCAAGCAGCGGCTGAAGCTGAAGGATGGCGGCCACTACGACGC
                     CGAGGTGAAAACCACCTACAAGGCCAAGAAACCCGTGCAGCTGCCTGGCGCCTACAAC
                     GTGAACATCAAGCTGGACATCACCAGCCACAACGAGGACTACACCATCGTGGAGCAGT
                     ACGAGCGGGCCGAGGGCAGACACAGCACCGGCGGCATGGACGAG"
                     /label="TACAAAGACGATGACGACAAGGGCAGCGTGTCCAAGGGCGAAGAGGACAA
                     CATGGCCATCATCAAGGAGTTCATGCGGTTCAAGGTGCACATGGAAGGCAGCGTGAAC
                     GGCCACGAGTTCGAGATCGAGGGCGAGGGCGAAGGCAGGCCCTACGAGGGCACCCAGA
                     CCGCCAAGCTGAAGGTGACCAAGGGCGGACCTCTGCCCTTCGCCTGGGACATCCTGAG
                     CCCCCAGTTCATGTACGGCAGCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGAC
                     TACCTGAAGCTGTCCTTCCCCGAGGGATTCAAATGGGAGCGGGTGATGAACTTCGAGG
                     ACGGCGGCGTGGTGACCGTGACCCAGGACAGCAGCCTGCAGGACGGCGAGTTCATCTA
                     CAAGGTGAAACTGCGGGGCACCAACTTCCCCAGCGACGGCCCCGTGATGCAGAAAAAG
                     ACCATGGGCTGGGAGGCCAGCAGCGAGCGGATGTACCCCGAGGATGGCGCCCTGAAGG
                     GCGAGATCAAGCAGCGGCTGAAGCTGAAGGATGGCGGCCACTACGACGCCGAGGTGAA
                     AACCACCTACAAGGCCAAGAAACCCGTGCAGCTGCCTGGCGCCTACAACGTGAACATC
                     AAGCTGGACATCACCAGCCACAACGAGGACTACACCATCGTGGAGCAGTACGAGCGGG
                     CCGAGGGCAGACACAGCACCGGCGGCATGGACGAG"
                     /note="Geneious type: Editing History Replacement"
     CDS             43..>951
                     /created_by="saibou"
                     /modified_by="saibou"
                     /label="CAHS3_DM_for_GeneArt"
     primer          complement(934..966)
                     /created_by="saibou"
                     /label="C3-stable2-rv"
     primer          952..971
                     /created_by="saibou"
                     /label="STABLE2-iR"
     misc_feature    962..969
                     /Recognition_pattern="GC^GGCCGC"
                     /Methylation="Mixed Methylation"
                     /label="NotI, CciNI"
                     /note="Geneious type: restriction site"
     CDS             970..1023
                     /note="<html><body>Eukaryotic ribosomes fail to insert a
                     peptide bond between the Gly and Pro residues, yielding
                     separate polypeptides.</body></html>"
                     /protein_id="<html><body><br></body></html>"
                     /transl_table=1
                     /codon_start=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /product="<html><body>2A peptide from&nbsp;<i>Thosea
                     asigna</i> virus capsid protein</body></html>"
                     /label="T2A"
     misc_feature    1030..1035
                     /Recognition_pattern="GAT^ATC"
                     /Methylation="Mixed Methylation"
                     /label="EcoRV, Eco32I"
                     /note="Geneious type: restriction site"
     misc_feature    1036..1041
                     /Recognition_pattern="T^CTAGA"
                     /Methylation="Dam:blocked"
                     /label="XbaI"
                     /note="Geneious type: restriction site"
     regulatory      1042..1051
                     /regulatory_class="other"
                     /note="<html><body>vertebrate consensus sequence for
                     strong initiation of translation (<a
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/3313277""
                     title=""https://www.ncbi.nlm.nih.gov/pubmed/3313277"">Koza
                     k, 1987</a>)</body></html>"
                     /label="Kozak sequence"
     CDS             join(1048..1050,1051..1053,1054..1764)
                     /note="<html><body>mammalian
                     codon-optimized</body></html>"
                     /transl_table=1
                     /codon_start=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK
                     LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIF
                     FKGDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQ
                     KNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKR
                     DHMVLLEFVTAAGITLGMDELYK"
                     /product="<html><body>the original enhanced GFP (<a
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/8948654""
                     title=""https://www.ncbi.nlm.nih.gov/pubmed/8948654"">Yang
                     et al., 1996</a>)</body></html>"
                     /label="EGFP"
     primer_bind     complement(1093..1114)
                     /Tm=67
                     /Description="EGFP, reverse primer"
                     /label="EGFP-N"
     primer          complement(1109..1128)
                     /created_by="saibou"
                     /modified_by="saibou"
                     /label="EGFP-seq"
     misc_feature    1328..1334
                     /Recognition_pattern="T^CCNGGA"
                     /label="PfoI"
                     /note="Geneious type: restriction site"
     primer_bind     1701..1722
                     /Tm=61
                     /Description="EGFP, forward primer"
                     /label="EGFP-C"
     misc_feature    1762..1767
                     /Recognition_pattern="A^AGCTT"
                     /Methylation="EcoBI:some blocked"
                     /label="HindIII"
                     /note="Geneious type: restriction site"
     CDS             1768..1821
                     /note="<html><body>Eukaryotic ribosomes fail to insert a
                     peptide bond between the Gly and Pro residues, yielding
                     separate polypeptides.</body></html>"
                     /protein_id="<html><body><br></body></html>"
                     /transl_table=1
                     /codon_start=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /product="<html><body>2A peptide from&nbsp;<i>Thosea
                     asigna</i> virus capsid protein</body></html>"
                     /label="T2A"
     misc_feature    1798..1803
                     /Recognition_pattern="CACGTC(-3/-3)"
                     /Methylation="Mixed Methylation"
                     /label="BtrI, AjiI, BmgBI"
                     /note="Geneious type: restriction site"
     misc_feature    1822..1827
                     /Recognition_pattern="G^CTAGC"
                     /Methylation="Mixed Methylation"
                     /label="NheI, AsuNHI"
                     /note="Geneious type: restriction site"
     misc_feature    1822..1827
                     /Recognition_pattern="GCTAG^C"
                     /Methylation="Mixed Methylation"
                     /label="BmtI, BspOI"
                     /note="Geneious type: restriction site"
     CDS             1828..2631
                     /note="<html><body>confers resistance to neomycin,
                     kanamycin, and G418 (Geneticin®)</body></html>"
                     /transl_table=1
                     /codon_start=1
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA
                     QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG
                     QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD
                     LDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA
                     DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /gene="<html><body><i>aph(3')-II</i> (or
                     <i>nptII</i>)</body></html>"
                     /label="NeoR/KanR"
     primer_bind     complement(1891..1910)
                     /Tm=57
                     /Description="Neomycin resistance gene, reverse primer"
                     /label="Neo-R"
     misc_feature    1986..1997
                     /Recognition_pattern="GACNNNN^NNGTC"
                     /Methylation="Mixed Methylation"
                     /label="DrdI, AasI, DseDI"
                     /note="Geneious type: restriction site"
     misc_feature    2080..2088
                     /Recognition_pattern="GACN^NNGTC"
                     /Methylation="Mixed Methylation"
                     /label="Tth111I, PflFI, PsyI"
                     /note="Geneious type: restriction site"
     misc_feature    2267..2272
                     /Recognition_pattern="YAC^GTR"
                     /Methylation="Mixed Methylation"
                     /label="BsaAI, BstBAI, Ppu21I"
                     /note="Geneious type: restriction site"
     misc_feature    2326..2331
                     /Recognition_pattern="GRGCY^C"
                     /Methylation="Mixed Methylation"
                     /label="BanII, Eco24I, EcoT38I, FriOI"
                     /note="Geneious type: restriction site"
     misc_feature    2366..2371
                     /Recognition_pattern="GCATG^C"
                     /Methylation="Mixed Methylation"
                     /label="SphI, PaeI"
                     /note="Geneious type: restriction site"
     misc_feature    2480..2486
                     /Recognition_pattern="CG^GWCCG"
                     /Methylation="Mixed Methylation"
                     /label="RsrII, CpoI, CspI, Rsr2I"
                     /note="Geneious type: restriction site"
     primer_bind     2501..2520
                     /Tm=57
                     /Description="Neomycin resistance gene, forward primer"
                     /label="Neo-F"
     misc_feature    2631..2638
                     /Recognition_pattern="VC^TCGAGB"
                     /Methylation="CpG:some impaired, EcoBI:cut, EcoKI:cut"
                     /label="PspXI"
                     /note="Geneious type: restriction site"
     misc_feature    2632..2637
                     /Recognition_pattern="C^TCGAG"
                     /Methylation="Mixed Methylation"
                     /label="XhoI, PaeR7I, Sfr274I, SlaI, StrI"
                     /note="Geneious type: restriction site"
     misc_feature    2638..2643
                     /Recognition_pattern="G^GATCC"
                     /Methylation="Dam:cut, Dcm:cut, CpG:cut"
                     /label="BamHI"
                     /note="Geneious type: restriction site"
     polyA_signal    2639..2773
                     /note="<html><body>SV40 polyadenylation
                     signal</body></html>"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(2663..2682)
                     /Tm=54
                     /Description="SV40 polyA terminator, reverse primer"
                     /label="EBV-rev"
     primer_bind     2717..2736
                     /Tm=51
                     /Description="SV40 polyA, reverse primer"
                     /label="SV40pA-R"
     misc_feature    2771..2776
                     /Recognition_pattern="GTT^AAC"
                     /Methylation="Mixed Methylation"
                     /label="HpaI, KspAI"
                     /note="Geneious type: restriction site"
     misc_feature    2782..2787
                     /Recognition_pattern="C^AATTG"
                     /Methylation="Mixed Methylation"
                     /label="MfeI, MunI"
                     /note="Geneious type: restriction site"
     misc_feature    2870..2879
                     /Recognition_pattern="GATNN^NNATC"
                     /Methylation="Mixed Methylation"
                     /label="BsaBI, Bse8I, BseJI"
                     /note="Geneious type: restriction site"
     misc_feature    2875..2880
                     /Recognition_pattern="T^GATCA"
                     /Methylation="Mixed Methylation"
                     /label="BclI, FbaI, Ksp22I"
                     /note="Geneious type: restriction site"
     misc_feature    2881..2886
                     /Recognition_pattern="GT^MKAC"
                     /Methylation="Mixed Methylation"
                     /label="AccI, FblI, XmiI"
                     /note="Geneious type: restriction site"
     misc_feature    2881..2886
                     /Recognition_pattern="G^TCGAC"
                     /Methylation="CpG:some blocked"
                     /label="SalI"
                     /note="Geneious type: restriction site"
     misc_feature    2942..2951
                     /Recognition_pattern="GACNN^NNGTC"
                     /Methylation="Mixed Methylation"
                     /label="PshAI, BoxI, BstPAI"
                     /note="Geneious type: restriction site"
     misc_feature    3005..3010
                     /Recognition_pattern="AGC^GCT"
                     /Methylation="Mixed Methylation"
                     /label="Eco47III, AfeI, Aor51HI"
                     /note="Geneious type: restriction site"
     misc_feature    3159..3169
                     /Recognition_pattern="GCCNNNN^NGGC"
                     /Methylation="Dcm:cut, CpG:variable"
                     /label="BglI"
                     /note="Geneious type: restriction site"
     misc_feature    3202..3207
                     /Recognition_pattern="TCG^CGA"
                     /Methylation="Mixed Methylation"
                     /label="NruI, Bsp68I, BtuMI, RruI"
                     /note="Geneious type: restriction site"
     misc_feature    3256..3267
                     /Recognition_pattern="(10/12)GCANNNNNNTGC(12/10)"
                     /label="AlfI"
                     /note="Geneious type: restriction site"
     misc_feature    3275..3285
                     /Recognition_pattern="GCANNNN^NTGC"
                     /Methylation="CpG:variable, EcoBI:blocked"
                     /label="BstAPI"
                     /note="Geneious type: restriction site"
     rep_origin      complement(3395..3983)
                     /note="<html><body>high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of replication</body></html>"
                     /direction="LEFT"
                     /label="ori"
     misc_feature    3436..3447
                     /Recognition_pattern="GACNNNN^NNGTC"
                     /Methylation="Mixed Methylation"
                     /label="DrdI, AasI, DseDI"
                     /note="Geneious type: restriction site"
     primer_bind     complement(3475..3494)
                     /Tm=56
                     /Description="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     CDS             join(complement(4946..5014),complement(4154..4945))
                     /note="<html><body>confers resistance to ampicillin,
                     carbenicillin, and related antibiotics</body></html>"
                     /transl_table=1
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVG
                     YIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLV
                     EYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
                     LDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGP
                     LLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQI
                     AEIGASLIKHW*"
                     /product="<html><body>β-lactamase</body></html>"
                     /gene="<html><body><i>bla</i></body></html>"
                     /label="AmpR"
     misc_feature    4222..4232
                     /Recognition_pattern="GACNNN^NNGTC"
                     /Methylation="Mixed Methylation"
                     /label="Eam1105I, AhdI, BmeRI, DriI"
                     /note="Geneious type: restriction site"
     misc_feature    4294..4299
                     /Recognition_pattern="GGTCTC(1/5)"
                     /Methylation="Mixed Methylation"
                     /label="Eco31I, BsaI, Bso31I"
                     /note="Geneious type: restriction site"
     misc_feature    4341..4351
                     /Recognition_pattern="GCCNNNN^NGGC"
                     /Methylation="Dcm:cut, CpG:variable"
                     /label="BglI"
                     /note="Geneious type: restriction site"
     misc_feature    4398..4403
                     /Recognition_pattern="AT^TAAT"
                     /Methylation="Mixed Methylation"
                     /label="VspI, AseI, PshBI"
                     /note="Geneious type: restriction site"
     misc_feature    4468..4473
                     /Recognition_pattern="CTGCA^G"
                     /label="PstI"
                     /note="Geneious type: restriction site"
     misc_feature    4594..4599
                     /Recognition_pattern="CGAT^CG"
                     /Methylation="Mixed Methylation"
                     /label="PvuI, BpvUI, MvrI, Ple19I"
                     /note="Geneious type: restriction site"
     misc_feature    4705..4710
                     /Recognition_pattern="AGT^ACT"
                     /Methylation="Mixed Methylation"
                     /label="ScaI, AssI, BmcAI, ZrmI"
                     /note="Geneious type: restriction site"
     primer_bind     4777..4796
                     /Tm=58
                     /Description="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     promoter        complement(5015..5119)
                     /gene="<html><body><i>bla</i></body></html>"
                     /label="AmpR promoter"
     primer_bind     5187..5205
                     /Tm=57
                     /Description="pBR322 vectors, upsteam of EcoRI site,
                     forward primer"
                     /label="pBRforEco"
     misc_feature    5223..5228
                     /Recognition_pattern="AT^TAAT"
                     /Methylation="Mixed Methylation"
                     /label="VspI, AseI, PshBI"
                     /note="Geneious type: restriction site"
     promoter        5269..7781
                     /note="<html><body><i>Drosophila
                     melanogaster</i>&nbsp;actin 5C promoter</body></html>"
                     /label="Ac5 promoter"
     misc_feature    5345..5350
                     /Recognition_pattern="A^GATCT"
                     /Methylation="Dam:cut, EcoBI:some blocked"
                     /label="BglII"
                     /note="Geneious type: restriction site"
     misc_feature    5590..5600
                     /Recognition_pattern="CCANNNN^NTGG"
                     /Methylation="Mixed Methylation"
                     /label="PflMI, AccB7I, BasI, Van91I"
                     /note="Geneious type: restriction site"
     misc_feature    5600..5607
                     /Recognition_pattern="GCGAT^CGC"
                     /Methylation="Mixed Methylation"
                     /label="SgfI, AsiSI, RgaI, SfaAI"
                     /note="Geneious type: restriction site"
     misc_feature    5601..5606
                     /Recognition_pattern="CGAT^CG"
                     /Methylation="Mixed Methylation"
                     /label="PvuI, BpvUI, MvrI, Ple19I"
                     /note="Geneious type: restriction site"
     misc_feature    5638..5643
                     /Recognition_pattern="C^CCGGG"
                     /Methylation="Mixed Methylation"
                     /label="Cfr9I, TspMI, XmaI"
                     /note="Geneious type: restriction site"
     misc_feature    5638..5643
                     /Recognition_pattern="CCC^GGG"
                     /Methylation="CpG:blocked"
                     /label="SmaI"
                     /note="Geneious type: restriction site"
     misc_feature    5714..5720
                     /Recognition_pattern="CCTCAGC(-5/-2)"
                     /Methylation="CpG:impaired, EcoBI:cut"
                     /label="BbvCI"
                     /note="Geneious type: restriction site"
     misc_feature    5725..5730
                     /Recognition_pattern="TCG^CGA"
                     /Methylation="Mixed Methylation"
                     /label="NruI, Bsp68I, BtuMI, RruI"
                     /note="Geneious type: restriction site"
     misc_feature    5808..5813
                     /Recognition_pattern="A^CATGT"
                     /Methylation="Mixed Methylation"
                     /label="PciI, PscI"
                     /note="Geneious type: restriction site"
     misc_feature    6309..6314
                     /Recognition_pattern="C^GTACG"
                     /Methylation="Mixed Methylation"
                     /label="BsiWI, Pfl23II, PspLI"
                     /note="Geneious type: restriction site"
     misc_feature    6314..6319
                     /Recognition_pattern="GCATG^C"
                     /Methylation="Mixed Methylation"
                     /label="SphI, PaeI"
                     /note="Geneious type: restriction site"
     misc_feature    6421..6426
                     /Recognition_pattern="GT^MKAC"
                     /Methylation="Mixed Methylation"
                     /label="AccI, FblI, XmiI"
                     /note="Geneious type: restriction site"
     misc_feature    6515..6526
                     /Recognition_pattern="(10/12)GCANNNNNNTGC(12/10)"
                     /label="AlfI"
                     /note="Geneious type: restriction site"
     misc_feature    6615..6620
                     /Recognition_pattern="ATGCA^T"
                     /Methylation="Mixed Methylation"
                     /label="EcoT22I, Mph1103I, NsiI, Zsp2I"
                     /note="Geneious type: restriction site"
     misc_feature    6630..6635
                     /Recognition_pattern="C^AATTG"
                     /Methylation="Mixed Methylation"
                     /label="MfeI, MunI"
                     /note="Geneious type: restriction site"
     misc_feature    6900..6910
                     /Recognition_pattern="GCANNNN^NTGC"
                     /Methylation="CpG:variable, EcoBI:blocked"
                     /label="BstAPI"
                     /note="Geneious type: restriction site"
     misc_feature    6920..6925
                     /Recognition_pattern="AT^CGAT"
                     /Methylation="Mixed Methylation"
                     /label="ClaI, Bsa29I, BseCI, BshVI, BspDI, Bsu15I"
                     /note="Geneious type: restriction site"
     misc_feature    6978..6987
                     /Recognition_pattern="GATNN^NNATC"
                     /Methylation="Mixed Methylation"
                     /label="BsaBI, Bse8I, BseJI"
                     /note="Geneious type: restriction site"
     misc_feature    7033..7038
                     /Recognition_pattern="T^GATCA"
                     /Methylation="Mixed Methylation"
                     /label="BclI, FbaI, Ksp22I"
                     /note="Geneious type: restriction site"
     misc_feature    7147..7152
                     /Recognition_pattern="C^TTAAG"
                     /label="AflII, BfrI, BspTI, BstAFI, MspCI, Vha464I"
                     /note="Geneious type: restriction site"
     misc_feature    7171..7176
                     /Recognition_pattern="A^CTAGT"
                     /Methylation="Mixed Methylation"
                     /label="SpeI, AhlI, BcuI"
                     /note="Geneious type: restriction site"
     misc_feature    7325..7330
                     /Recognition_pattern="A^CATGT"
                     /Methylation="Mixed Methylation"
                     /label="PciI, PscI"
                     /note="Geneious type: restriction site"
     misc_feature    7332..7337
                     /Recognition_pattern="CA^TATG"
                     /Methylation="Mixed Methylation"
                     /label="NdeI, FauNDI"
                     /note="Geneious type: restriction site"
     misc_feature    7469..7475
                     /Recognition_pattern="RG^GWCCY"
                     /Methylation="Mixed Methylation"
                     /label="PpuMI, Psp5II, PspPPI"
                     /note="Geneious type: restriction site"
     misc_feature    7571..7583
                     /Recognition_pattern="(7/12)GAACNNNNNNTAC(12/7)"
                     /Methylation="CpG:impaired"
                     /label="PsrI"
                     /note="Geneious type: restriction site"
     misc_feature    7648..7653
                     /Recognition_pattern="AGC^GCT"
                     /Methylation="Mixed Methylation"
                     /label="Eco47III, AfeI, Aor51HI"
                     /note="Geneious type: restriction site"
     misc_feature    7651..7657
                     /Recognition_pattern="GC^TNAGC"
                     /Methylation="Mixed Methylation"
                     /label="BlpI, Bpu1102I, Bsp1720I"
                     /note="Geneious type: restriction site"
     primer          7737..7757
                     /created_by="saibou"
                     /modified_by="saibou"
                     /label="Ac5 forward primer"
     misc_feature    7777..7782
                     /Recognition_pattern="GGTCTC(1/5)"
                     /Methylation="Mixed Methylation"
                     /label="Eco31I, BsaI, Bso31I"
                     /note="Geneious type: restriction site"
ORIGIN      
        1 ggtacctact agtccagtgt ggtggaattc gccaccatgg acatgagtag ccgccagaat
       61 cagcagagca gcagccagca tagtagcagc tcccagcaag gcggacaagg tggacagggt
      121 gttcaaggca gcagcagcta tagccgcacc gaggttcaca caagcagcgg cggaccaaca
      181 attggaggtg cccaacgtac agtgccagtg ccaccaggat cgcatagcga ggtgcacgaa
      241 gaacgcgaag tgatcaagca cggcaccaag accgagagcg agacacacgt tgtgaccgtg
      301 ccagtgacaa ccttcggcag caccaacatg gaaagcgtgc gcaccggatt caccgtgacg
      361 caggataaga acctgaccgt ggccgctccg aatatcgccg ctccaatcca cagcaacctg
      421 gatctgaatc tcggaggcgg agcccgtgcc gaaattaccg ctggaacaac ggtggatctg
      481 agcaagatcc agcgcaagga tctgggcccc gaagagtacg cccgttacaa ggccaaggtg
      541 gaacagctgg cccgccagga tgaacaggat gctggaatgc gtgccgcgca gtaccgtgaa
      601 gaggtggaac gtgatgccga gctgatccgc cagattctgg aacgccagca catccgcgat
      661 ttggagttcc gcaaagagat ggtcgagaat caagtgaacc gccaagagcg cgagatccag
      721 ttggaggccg aatatgccat gcgcgccctg gaactggaac gcaatgccgc caaagaagcc
      781 ctggaaagcg ccaaggctca gaccaacgtg aacgtgaagg tcgagagcgc catcggaacc
      841 acagtgtcca agggcgccat tcagaccagc gccgataaga gcagcaccac caagacagga
      901 cccaccaccg tgacacagat taagcacacc gagcagcata ccgagcgccg cctgtataaa
      961 ggcggccgcg aaggacgcgg cagcctactg acttgcggag atgtcgaaga gaaccctggc
     1021 cctggttccg atatctctag agccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1081 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1141 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1201 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1261 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1321 ggctacgtcc aggagcgcac catcttcttc aagggcgacg gcaactacaa gacccgcgcc
     1381 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1441 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1501 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1561 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1621 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1681 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1741 ctcggcatgg acgagctgta caagcttgag ggcagaggaa gtcttctaac atgcggtgac
     1801 gtggaggaga atcccggccc tgctagcatg ggatcggcca ttgaacaaga tggattgcac
     1861 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     1921 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     1981 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     2041 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     2101 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     2161 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     2221 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     2281 gaagccggtc ttgtcgatca ggatgatctg gacgaagaac atcaggggct cgcgccagcc
     2341 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     2401 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     2461 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     2521 gctgaagaac ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     2581 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg actcgaggga
     2641 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2701 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2761 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2821 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     2881 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2941 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     3001 cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg
     3061 gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc
     3121 ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc
     3181 tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc
     3241 ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag
     3301 atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc cagcaaaagg
     3361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     3601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4441 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     4501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4741 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     4801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5161 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     5221 gaattaattc tatattctaa aaacacaaat gatacttcta aaaaaaatca tgaatggcat
     5281 caactctgaa tcaaatcttt gcagatgcac ctacttctca tttccactgt cacatcattt
     5341 ttccagatct cgctgcctgt tatgtggccc acaaaccaag acacgtttta tggccattaa
     5401 agctggctga tcgtcgccaa acaccaaata catatcaata tgtacattcg agaaagaagc
     5461 gatcaaagaa gcgtcttcgg gcgagtagga gaatgcggag gagaaggaga acgagctgat
     5521 ctagtatctc tccacaatcc aatgccaact gaccaactgg ccatattcgg agcaatttga
     5581 agccaatttc catcgcctgg cgatcgctcc attcttggct atatgttttt caccgttccc
     5641 ggggccattt tcaaagactc gtcggtaaga taagattgtg tcactcgctg tctctcttca
     5701 tttgtcgaag aatgctgagg aatttcgcga tgacgtcggc gagtattttg aagaatgaga
     5761 ataatttgta tttatacgaa aatcagttag tggaattttc tacaaaaaca tgttatctat
     5821 agataatttt gttgcaaaat atgttgacta tgacaaagat tgtatgtata tacctttaat
     5881 gtattctcat tttcttatgt atttataatg gcaatgatga tactgatgat attttaagat
     5941 gatgccagac cacaggctga tttctgcgtc ttttgccgaa cgcagtgcat gtgcggttgt
     6001 tgttttttgg aatagtttca attttcggac tgtccgcttt gatttcagtt tcttggctta
     6061 ttcaaaaagc aaagtaaagc caaaaaagcg agatggcaat accaaatgcg gcaaaacggt
     6121 agtggaagga aaggggtgcg gggcagcgga aggaagggtg gggcggggcg tggcggggtc
     6181 tgtggctggg cgcgacgtca ccgacgttgg agccactcct ttgaccatgt gtgcgtgtgt
     6241 gtattattcg tgtctcgcca ctcgccggtt gtttttttct ttttatctcg ctctctctag
     6301 cgccatctcg tacgcatgct caacgcaccg catgttgccg tgtcctttat gcgtcatttt
     6361 ggctcgaaat aggcaattat ttaaacaaag attagtcaac gaaaacgcta aaataaataa
     6421 gtctacaata tggttactta ttgccatgtg tgtgcagcca acgatagcaa caaaagcaac
     6481 aacacagtgg ctttccctct ttcacttttt gtttgcaagc gcgtgcgagc aagacggcac
     6541 gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag ttttggccga aaaacatcaa
     6601 ggcgcctgat acgaatgcat ttgcaataac aattgcgata tttaatattg tttatgaagc
     6661 tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa ttatttgaaa gagaattagg
     6721 aatcggacag cttatcgtta cgggctaaca gcacaccgag acgaaatagc ttacctgacg
     6781 tcacagcctc tggaagaact gccgccaagc agagagagag agaaaaagag ggagagcagc
     6841 ttagaccgca tgtgcttgtg tgtgaggcgt ctctctcttc gtctcctgtt tgcgcaaacg
     6901 catagactgc actgagaaaa tcgattacct attttttatg aatgaatatt tgcactatta
     6961 ctattcaaaa ctattaagat agcaatcaca ttcaatagcc aaatactata ccacctgagc
     7021 gatgcaacga aatgatcaat ttgagcaaaa atgctgcata tttaggacgg catcattata
     7081 gaaatgcttc ttgctgtgta cttttctctc gtctggcagc tgtttcgccg ttattgttaa
     7141 aaccggctta agttaggtgt gttttctacg actagtgatg cccctactag aagatgtgtg
     7201 ttgcacaaat gtccctgaat aaccaatttg aagtgcagat agcagtaaac gtaagctaat
     7261 atgaatatta tttaactgta atgttttaat atcgctggac attactaata aacccactat
     7321 aaacacatgt acatatgtat gttttggcat acaatgagta gttggggaaa aaatgtgtaa
     7381 aagcaccgtg accatcacag cataaagata accagctgaa gtatcgaata tgagtaaccc
     7441 ccaaattgaa tcacatgccg caactgatag gacccatgga agtacactct tcatggcgat
     7501 atacaagaca cacacaagca cgaacaccca gttgcggagg aaattctccg taaatgaaaa
     7561 cccaatcggc gaacaattca tacccatata tggtaaaagt tttgaacgcg acttgagagc
     7621 ggagagcatt gcggctgata aggttttagc gctaagcggg ctttataaaa cgggctgcgg
     7681 gaccagtttt catatcacta ccgtttgagt tcttgtgctg tgtggatact cctcccgaca
     7741 caaagccgct ccatcagcca gcagtcgtct aatccagaga ccccggatcg g
//
