LOCUS       EML8357_pA3A-CBE       18063 bp    DNA     circular SYN 31-MAR-2025
DEFINITION  CRISPR/Cas9-mediated base editing in Arabidopsis. Contains 
            Cas9(D10A) and empty gRNA scaffold, HygR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18063)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 31, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..18063
                     /mol_type="genomic DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /locus_tag="RB T-DNA repeat"
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
                     /note="/ApEinfo_fwdcolor=#7eff74"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=RB T-DNA repeat"
                     /note="/ApEinfo_revcolor=#7eff74"
     primer_bind     213..235
                     /locus_tag="M13/pUC Forward"
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=M13/pUC Forward"
                     /note="/ApEinfo_revcolor=#4ec02b"
     primer_bind     227..244
                     /locus_tag="M13 Forward"
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=M13 Forward"
                     /note="/ApEinfo_revcolor=#4ec02b"
     primer_bind     228..244
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=M13 fwd"
                     /note="/ApEinfo_revcolor=#4ec02b"
     CDS             957..1748
                     /codon_start=1
                     /gene="aadA"
                     /locus_tag="SmR"
                     /product="aminoglycoside adenylyltransferase (Murphy,
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and
                     streptomycin"
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=SmR"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        1900..1975
                     /locus_tag="gRNA scaffold"
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=gRNA scaffold"
                     /note="/ApEinfo_revcolor=pink"
     misc_feature    2184..3535
                     /locus_tag="ECp"
                     /label=ECp
                     /note="/ApEinfo_fwdcolor=#80ff80"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=ECp"
                     /note="/ApEinfo_revcolor=#80ff80"
     misc_feature    2184..3535
                     /locus_tag="ECp"
                     /label=ECp copy
                     /label=ECp
                     /note="/ApEinfo_fwdcolor=#80ff80"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=ECp"
                     /note="/ApEinfo_revcolor=#80ff80"
     misc_feature    3542..4192
                     /locus_tag="A3A/Y130F"
                     /label=A3A/Y130F
                     /note="/ApEinfo_fwdcolor=#0080ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=A3A/Y130F"
                     /note="/ApEinfo_revcolor=#0080ff"
     misc_feature    3542..4192
                     /locus_tag="A3A/Y130F"
                     /label=A3A/Y130F copy
                     /label=A3A/Y130F
                     /note="/ApEinfo_fwdcolor=#0080ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=A3A/Y130F"
                     /note="/ApEinfo_revcolor=#0080ff"
     misc_feature    4153
                     /locus_tag="removeBsaI GAGACC-GAGACT"
                     /label=removeBsaI GAGACC-GAGACT
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=removeBsaI GAGACC-GAGACT"
                     /note="/ApEinfo_revcolor=green"
     CDS             4193..4258
                     /codon_start=1
                     /locus_tag="3xFLAG"
                     /product="three tandem FLAG(R) epitope tags, followed by
                     an enterokinase cleavage site"
                     /label=3xFLAG
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=3xFLAG"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4265..4285
                     /codon_start=1
                     /locus_tag="SV40 NLS"
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /label=SV40 NLS
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=SV40 NLS"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="PKKKRKV"
     CDS             4310..8410
                     /codon_start=1
                     /locus_tag="Cas9(D10A)"
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due
                     to the D10A mutation in the RuvC catalytic domain"
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=Cas9(D10A)"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8323..8331
                     /locus_tag="Shine-Dalgarno sequence"
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=Shine-Dalgarno sequence"
                     /note="/ApEinfo_revcolor=cyan"
     CDS             8411..8458
                     /codon_start=1
                     /locus_tag="nucleoplasmin NLS"
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=nucleoplasmin NLS"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8480..8728
                     /codon_start=1
                     /locus_tag="UGI"
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=UGI"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8741..8761
                     /codon_start=1
                     /locus_tag="SV40 NLS(1)"
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /label=SV40 NLS(1)
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=SV40 NLS"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="PKKKRKV"
     misc_feature    8765..9403
                     /locus_tag="rbsT"
                     /label=rbsT
                     /note="/ApEinfo_fwdcolor=#800000"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=rbsT"
                     /note="/ApEinfo_revcolor=#800000"
     misc_feature    8765..9403
                     /locus_tag="rbsT"
                     /label=rbsT copy
                     /label=rbsT
                     /note="/ApEinfo_fwdcolor=#800000"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=rbsT"
                     /note="/ApEinfo_revcolor=#800000"
     primer_bind     complement(9418..9434)
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=M13 rev"
                     /note="/ApEinfo_revcolor=#4ec02b"
     primer_bind     complement(9418..9434)
                     /locus_tag="M13 Reverse"
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=M13 Reverse"
                     /note="/ApEinfo_revcolor=#4ec02b"
     primer_bind     complement(9431..9453)
                     /locus_tag="M13/pUC Reverse"
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=M13/pUC Reverse"
                     /note="/ApEinfo_revcolor=#4ec02b"
     protein_bind    9442..9458
                     /locus_tag="lac operator"
                     /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)."
                     /note="/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=lac operator"
                     /note="/ApEinfo_revcolor=pink"
     promoter        complement(9466..9496)
                     /locus_tag="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="/ApEinfo_fwdcolor=#346ee0"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=lac promoter"
                     /note="/ApEinfo_revcolor=#346ee0"
     protein_bind    9511..9532
                     /locus_tag="CAP binding site"
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /note="/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=CAP binding site"
                     /note="/ApEinfo_revcolor=pink"
     promoter        9723..10399
                     /locus_tag="CaMV 35S promoter (enhanced)"
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
                     /note="/ApEinfo_fwdcolor=#346ee0"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=CaMV 35S promoter (enhanced)"
                     /note="/ApEinfo_revcolor=#346ee0"
     promoter        9723..10399
                     /locus_tag="CaMV 35S promoter (enhanced)"
                     /label=CaMV 35S promoter (enhanced) copy
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
                     /note="/ApEinfo_fwdcolor=#346ee0"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=CaMV 35S promoter (enhanced)"
                     /note="/ApEinfo_revcolor=#346ee0"
     primer_bind     10336..10356
                     /locus_tag="35S promoter"
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=35S promoter"
                     /note="/ApEinfo_revcolor=#4ec02b"
     CDS             10466..11491
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /locus_tag="HygR"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=HygR"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    11532..11706
                     /locus_tag="CaMV poly(A) signal"
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
                     /note="/ApEinfo_fwdcolor=#ff3eee"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=CaMV poly(A) signal"
                     /note="/ApEinfo_revcolor=#ff3eee"
     misc_feature    11784..11808
                     /locus_tag="LB T-DNA repeat"
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
                     /note="/ApEinfo_fwdcolor=#7eff74"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=LB T-DNA repeat"
                     /note="/ApEinfo_revcolor=#7eff74"
     CDS             12233..13027
                     /codon_start=1
                     /gene="aphA-3"
                     /locus_tag="KanR"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=KanR"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      13114..13702
                     /direction=RIGHT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="/ApEinfo_fwdcolor=#999999"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=ori"
                     /note="/ApEinfo_revcolor=#999999"
     primer_bind     13603..13622
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=pBR322ori-F"
                     /note="/ApEinfo_revcolor=#4ec02b"
     primer_bind     13856..13873
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=L4440"
                     /note="/ApEinfo_revcolor=#4ec02b"
     misc_feature    13888..14028
                     /locus_tag="bom"
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="/ApEinfo_fwdcolor=#7eff74"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=bom"
                     /note="/ApEinfo_revcolor=#7eff74"
     primer_bind     complement(13954..13973)
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=pRS-marker"
                     /note="/ApEinfo_revcolor=#4ec02b"
     primer_bind     14114..14136
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
                     /note="/ApEinfo_fwdcolor=#14c0bd"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=pGEX 3'"
                     /note="/ApEinfo_revcolor=#4ec02b"
     rep_origin      14372..14566
                     /locus_tag="pVS1 oriV"
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /note="/ApEinfo_fwdcolor=#999999"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=pVS1 oriV"
                     /note="/ApEinfo_revcolor=#999999"
     CDS             complement(14632..15705)
                     /codon_start=1
                     /locus_tag="pVS1 RepA"
                     /product="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=pVS1 RepA"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(16134..16763)
                     /codon_start=1
                     /locus_tag="pVS1 StaA"
                     /product="stability protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0}
                     width 5 offset 0"
                     /note="/ApEinfo_label=pVS1 StaA"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
BASE COUNT         4476 a         4550 c         4711 g         4326 t
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
      301 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
      361 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
      421 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
      481 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
      541 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
      601 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
      661 gagtcgaagt agtgattggg agaccaaccc agtggacata agcctgttcg gttcgtaagc
      721 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
      781 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
      841 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
      901 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatgg
      961 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     1021 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     1081 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     1141 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     1201 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     1261 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     1321 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     1381 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     1441 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     1501 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     1561 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     1621 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     1681 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     1741 gcaaataatg tctagctaga aattcgttca agccgacgcc gcttcgcggc gcggcttaac
     1801 tcaagcgtta gatgcactaa gcacataatt gctcacagcc aaactatcag gtcaagtctg
     1861 cttttattat ttttaagcgt gcataataag ccggtctcgg ttttagagct agaaatagca
     1921 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     1981 tttgcaaaat tttccagatc gatttcttct tcctctgttc ttcggcgttc aatttctggg
     2041 gttttctctt cgttttctgt aactgaaacc taaaatttga cctaaaaaaa atctcaaata
     2101 atatgattca gtggttttgt acttttcagt tagttgagtt ttgcagttcc gatgagataa
     2161 accaatacca tggttatact agtgaataaa agcatttgcg tttggtttat cattgcgttt
     2221 atacaaggac agagatccac tgagctggaa tagcttaaaa ccattatcag aacaaaataa
     2281 accatttttt gttaagaatc agagcatagt aaacaacaga aacaacctaa gagaggtaac
     2341 ttgtccaaga agatagctaa ttatatctat tttataaaag ttatcatagt ttgtaagtca
     2401 caaaagatgc aaataacaga gaaactagga gacttgagaa tatacattct tgtatatttg
     2461 tattcgagat tgtgaaaatt tgaccataag tttaaattct taaaaagata tatctgatct
     2521 aggtgatggt tatagactgt aattttacca catgtttaat gatggatagt gacacacatg
     2581 acacatcgac aacactatag catcttattt agattacaac atgaaatttt tctgtaatac
     2641 atgtctttgt acataattta aaagtaattc ctaagaaata tatttataca aggagtttaa
     2701 agaaaacata gcataaagtt caatgagtag taaaaaccat atacagtata tagcataaag
     2761 ttcaatgagt ttattacaaa agcattggtt cactttctgt aacacgacgt taaaccttcg
     2821 tctccaatag gagcgctact gattcaacat gccaatatat actaaatacg tttctacagt
     2881 caaatgcttt aacgtttcat gattaagtga ctatttaccg tcaatccttt cccattcctc
     2941 ccactaatcc aactttttaa ttactcttaa atcaccacta agctagtaac gcctatcatg
     3001 aattagctct actaaatcta gcaacctttc aaatttgcag tattgcaggt gtctctgtgt
     3061 ctttaaaata gttgccttat gatttcttcg gtttcaagat gatcaaatag ttatagattt
     3121 catgctcaca catgctcatt agatgtgtac atactttact tacccaaatc tattttctcg
     3181 caaagatttt gatggtaaag ctgatttggt tctattgaac taaatcaaac gagtttcaga
     3241 ctgagtgatt ctaatccggc ccattagccc ctaaacagac ccactaatta cgcagctttt
     3301 aatagagtaa ttacacctag tttacccact aaaccactaa gcactaatta tctcacaatc
     3361 taatgagctt ccctcgtaat tacttgggct ttcactctac catttatttg taacagtcaa
     3421 gtctctactg tctctatata aactctctaa agttaacaca caattctcat cacaaacaaa
     3481 tcaaccaaag caacttctac tctttcttct ttcgacctta tcaatctgtt gagaaatcta
     3541 gatggaggcc tctccggctt caggtcctcg ccatctcatg gacccccata tctttacctc
     3601 caatttcaat aacggcattg gcaggcacaa gacatacctc tgttatgagg tcgagcggct
     3661 cgataacgga acctcagtga agatggacca acacaggggt ttccttcata accaagcgaa
     3721 aaatctcctt tgcggtttct atgggcggca cgcggagctg agattcttgg atttggtccc
     3781 gagcctccag ctggatcccg cgcaaatata ccgcgtcact tggttcatat catggagccc
     3841 gtgctttagc tgggggtgtg ccggtgaggt tcgcgcgttc ctccaggaaa acacccatgt
     3901 tcgcctccgg attttcgcag cgaggatttt cgactatgat ccactttaca aagaggccct
     3961 ccaaatgctt agagatgctg gggctcaagt ttcaattatg acatacgacg agtttaagca
     4021 ttgttgggac acttttgtcg atcatcaggg ttgtcccttc cagccgtggg atggtcttga
     4081 tgagcattcg caagctttga gtggcaggct tcgcgcgata cttcagaacc agggtaactc
     4141 aggttctgag actcctggca caagtgagtc agcaacaccc gagtccacca tggattacaa
     4201 ggaccacgac ggggattaca aggaccacga cattgattac aaggatgatg atgacaagat
     4261 ggctccgaag aagaagagga aggttggcat ccacggggtg ccagctgctg acaagaagta
     4321 ctcgatcggc ctcgctattg ggactaactc tgttggctgg gccgtgatca ccgacgagta
     4381 caaggtgccc tcaaagaagt tcaaggtcct gggcaacacc gatcggcatt ccatcaagaa
     4441 gaatctcatt ggcgctctcc tgttcgacag cggcgagacg gctgaggcta cgcggctcaa
     4501 gcgcaccgcc cgcaggcggt acacgcgcag gaagaatcgc atctgctacc tgcaggagat
     4561 tttctccaac gagatggcga aggttgacga ttctttcttc cacaggctgg aggagtcatt
     4621 cctcgtggag gaggataaga agcacgagcg gcatccaatc ttcggcaaca ttgtcgacga
     4681 ggttgcctac cacgagaagt accctacgat ctaccatctg cggaagaagc tcgtggactc
     4741 cacagataag gcggacctcc gcctgatcta cctcgctctg gcccacatga ttaagttcag
     4801 gggccatttc ctgatcgagg gggatctcaa cccggacaat agcgatgttg acaagctgtt
     4861 catccagctc gtgcagacgt acaaccagct cttcgaggag aaccccatta atgcgtcagg
     4921 cgtcgacgcg aaggctatcc tgtccgctag gctctcgaag tctcggcgcc tcgagaacct
     4981 gatcgcccag ctgccgggcg agaagaagaa cggcctgttc gggaatctca ttgcgctcag
     5041 cctggggctc acgcccaact tcaagtcgaa tttcgatctc gctgaggacg ccaagctgca
     5101 gctctccaag gacacatacg acgatgacct ggataacctc ctggcccaga tcggcgatca
     5161 gtacgcggac ctgttcctcg ctgccaagaa tctgtcggac gccatcctcc tgtctgatat
     5221 tctcagggtg aacaccgaga ttacgaaggc tccgctctca gcctccatga tcaagcgcta
     5281 cgacgagcac catcaggatc tgaccctcct gaaggcgctg gtcaggcagc agctccccga
     5341 gaagtacaag gagatcttct tcgatcagtc gaagaacggc tacgctgggt acattgacgg
     5401 cggggcctct caggaggagt tctacaagtt catcaagccg attctggaga agatggacgg
     5461 cacggaggag ctgctggtga agctcaatcg cgaggacctc ctgaggaagc agcggacatt
     5521 cgataacggc agcatcccac accagattca tctcggggag ctgcacgcta tcctgaggag
     5581 gcaggaggac ttctaccctt tcctcaagga taaccgcgag aagatcgaga agattctgac
     5641 tttcaggatc ccgtactacg tcggcccact cgctaggggc aactcccgct tcgcttggat
     5701 gacccgcaag tcagaggaga cgatcacgcc gtggaacttc gaggaggtgg tcgacaaggg
     5761 cgctagcgct cagtcgttca tcgagaggat gacgaatttc gacaagaacc tgccaaatga
     5821 gaaggtgctc cctaagcact cgctcctgta cgagtacttc acagtctaca acgagctgac
     5881 taaggtgaag tatgtgaccg agggcatgag gaagccggct ttcctgtctg gggagcagaa
     5941 gaaggccatc gtggacctcc tgttcaagac caaccggaag gtcacggtta agcagctcaa
     6001 ggaggactac ttcaagaaga ttgagtgctt cgattcggtc gagatctctg gcgttgagga
     6061 ccgcttcaac gcctccctgg ggacctacca cgatctcctg aagatcatta aggataagga
     6121 cttcctggac aacgaggaga atgaggatat cctcgaggac attgtgctga cactcactct
     6181 gttcgaggac cgggagatga tcgaggagcg cctgaagact tacgcccatc tcttcgatga
     6241 caaggtcatg aagcagctca agaggaggag gtacaccggc tgggggaggc tgagcaggaa
     6301 gctcatcaac ggcattcggg acaagcagtc cgggaagacg atcctcgact tcctgaagag
     6361 cgatggcttc gcgaaccgca atttcatgca gctgattcac gatgacagcc tcacattcaa
     6421 ggaggatatc cagaaggctc aggtgagcgg ccagggggac tcgctgcacg agcatatcgc
     6481 gaacctcgct ggctcgccag ctatcaagaa ggggattctg cagaccgtga aggttgtgga
     6541 cgagctggtg aaggtcatgg gcaggcacaa gcctgagaac atcgtcattg agatggcccg
     6601 ggagaatcag accacgcaga agggccagaa gaactcacgc gagaggatga agaggatcga
     6661 ggagggcatt aaggagctgg ggtcccagat cctcaaggag cacccggtgg agaacacgca
     6721 gctgcagaat gagaagctct acctgtacta cctccagaat ggccgcgata tgtatgtgga
     6781 ccaggagctg gatattaaca ggctcagcga ttacgacgtc gatcatatcg ttccacagtc
     6841 attcctgaag gatgactcca ttgacaacaa ggtcctcacc aggtcggaca agaaccgggg
     6901 caagtctgat aatgttcctt cagaggaggt cgttaagaag atgaagaact actggcgcca
     6961 gctcctgaat gccaagctga tcacgcagcg gaagttcgat aacctcacaa aggctgagag
     7021 gggcgggctc tctgagctgg acaaggcggg cttcatcaag aggcagctgg tcgagacacg
     7081 gcagatcact aagcacgttg cgcagattct cgactcacgg atgaacacta agtacgatga
     7141 gaatgacaag ctgatccgcg aggtgaaggt catcaccctg aagtcaaagc tcgtctccga
     7201 cttcaggaag gatttccagt tctacaaggt tcgggagatc aacaattacc accatgccca
     7261 tgacgcgtac ctgaacgcgg tggtcggcac agctctgatc aagaagtacc caaagctcga
     7321 gagcgagttc gtgtacgggg actacaaggt ttacgatgtg aggaagatga tcgccaagtc
     7381 ggagcaggag attggcaagg ctaccgccaa gtacttcttc tactctaaca ttatgaattt
     7441 cttcaagaca gagatcactc tggccaatgg cgagatccgg aagcgccccc tcatcgagac
     7501 gaacggcgag acgggggaga tcgtgtggga caagggcagg gatttcgcga ccgtcaggaa
     7561 ggttctctcc atgccacaag tgaatatcgt caagaagaca gaggtccaga ctggcgggtt
     7621 ctctaaggag tcaattctgc ctaagcggaa cagcgacaag ctcatcgccc gcaagaagga
     7681 ctgggatccg aagaagtacg gcgggttcga cagccccact gtggcctact cggtcctggt
     7741 tgtggcgaag gttgagaagg gcaagtccaa gaagctcaag agcgtgaagg agctgctggg
     7801 gatcacgatt atggagcgct ccagcttcga gaagaacccg atcgatttcc tggaggcgaa
     7861 gggctacaag gaggtgaaga aggacctgat cattaagctc cccaagtact cactcttcga
     7921 gctggagaac ggcaggaagc ggatgctggc ttccgctggc gagctgcaga aggggaacga
     7981 gctggctctg ccgtccaagt atgtgaactt cctctacctg gcctcccact acgagaagct
     8041 caagggcagc cccgaggaca acgagcagaa gcagctgttc gtcgagcagc acaagcatta
     8101 cctcgacgag atcattgagc agatttccga gttctccaag cgcgtgatcc tggccgacgc
     8161 gaatctggat aaggtcctct ccgcgtacaa caagcaccgc gacaagccaa tcagggagca
     8221 ggctgagaat atcattcatc tcttcaccct gacgaacctc ggcgcccctg ctgctttcaa
     8281 gtacttcgac acaactatcg atcgcaagag gtacacaagc actaaggagg tcctggacgc
     8341 gaccctcatc caccagtcga ttaccggcct ctacgagacg cgcatcgacc tgtctcagct
     8401 cgggggcgac aagcggccag cggcgacgaa gaaggcgggg caggcgaaga agaagaaggg
     8461 agacggatca ggcgggtcaa caaacctcag tgacatcata gagaaggaaa ctggtaagca
     8521 actggttatt caagagagca ttctgatgct ccctgaggaa gtcgaggaag ttataggaaa
     8581 caagcctgag agcgatatac tcgtgcatac ggcgtatgac gagagcacgg atgaaaatgt
     8641 gatgctcttg accagcgatg cgccagaata taaaccctgg gcactggtta ttcaggactc
     8701 taacggggaa aataaaatca agatgctctc aggtggctcc ccaaagaaga aacgcaaggt
     8761 ttgagctcag agctttcgtt cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca
     8821 tcagtttcat tgcgcacaca ccagaatcct actgagtttg agtattatgg cattgggaaa
     8881 actgtttttc ttgtaccatt tgttgtgctt gtaatttact gtgtttttta ttcggttttc
     8941 gctatcgaac tgtgaaatgg aaatggatgg agaagagtta atgaatgata tggtcctttt
     9001 gttcattctc aaattaatat tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa
     9061 attataagag atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta
     9121 atgaccgaag ttaatatgag gagtaaaaca cttgtagttg taccattatg cttattcact
     9181 aggcaacaaa tatattttca gacctagaaa agctgcaaat gttactgaat acaagtatgt
     9241 cctcttgtgt tttagacatt tatgaacttt cctttatgta attttccaga atccttgtca
     9301 gattctaatc attgctttat aattatagtt atactcatgg atttgtagtt gagtatgaaa
     9361 atatttttta atgcatttta tgacttgcca attgattgac aacgaattcg taatcatgtc
     9421 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     9481 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     9541 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     9601 ccaacgcgcg gggagaggcg gtttgcgtat tggctagagc agcttgccaa catggtggag
     9661 cacgacactc tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct
     9721 attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct
     9781 atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat
     9841 tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga
     9901 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
     9961 gtggattgat gtgaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga
    10021 tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa
    10081 cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga
    10141 aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc
    10201 tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga
    10261 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga
    10321 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
    10381 tttggagagg acacgctgaa atcaccagtc tctctctaca aatctatctc tctcgagctt
    10441 tcgcagatcc ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga
    10501 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
    10561 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
    10621 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
    10681 cccgattccg gaagtgcttg acattgggga gtttagcgag agcctgacct attgcatctc
    10741 ccgccgttca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
    10801 acaaccggtc gcggaggcta tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
    10861 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
    10921 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
    10981 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
    11041 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
    11101 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
    11161 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
    11221 gcatccggag cttgcaggat cgccacgact ccgggcgtat atgctccgca ttggtcttga
    11281 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
    11341 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
    11401 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
    11461 ccccagcact cgtccgaggg caaagaaata gagtagatgc cgaccgggat ctgtcgatcg
    11521 acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt
    11581 cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
    11641 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc
    11701 cagatccccc gaattaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta
    11761 ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa
    11821 cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcagtc
    11881 cgggacggcg tcagcgggag agccgttgta aggcggcaga ctttgctcat gttaccgatg
    11941 ctattcggaa gaacggcaac taagctgccg ggtttgaaac acggatgatc tcgcggaggg
    12001 tagcatgttg attgtaacga tgacagagcg ttgctgcctg tgatcaccgc ggtttcaaaa
    12061 tcggctccgt cgatactatg ttatacgcca actttgaaaa caactttgaa aaagctgttt
    12121 tctggtattt aaggttttag aatgcaagga acagtgaatt ggagttcgtc ttgttataat
    12181 tagcttcttg gggtatcttt aaatactgta gaaaagagga aggaaataat aaatggctaa
    12241 aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac
    12301 ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt
    12361 aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat
    12421 gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga
    12481 tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga
    12541 agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca
    12601 ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt
    12661 ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg aagaagacac
    12721 tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga
    12781 acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt
    12841 aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt
    12901 ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga
    12961 cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt
    13021 gttttagtac ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
    13081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
    13141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
    13201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
    13261 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
    13321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
    13381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
    13441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
    13501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
    13561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
    13621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
    13681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
    13741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
    13801 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
    13861 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
    13921 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
    13981 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
    14041 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
    14101 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
    14161 gcgaggcagg gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc
    14221 gcgccctggt agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat
    14281 aggccgacgc gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg
    14341 cagctcttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt
    14401 tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt
    14461 cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg
    14521 gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa acagaccttt
    14581 tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc
    14641 ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg
    14701 ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac
    14761 ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa
    14821 caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc
    14881 gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc
    14941 ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc
    15001 ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac
    15061 caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg
    15121 aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt
    15181 gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg
    15241 gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca
    15301 cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta
    15361 gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat
    15421 gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg
    15481 ctcgtcgccc ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga
    15541 ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc
    15601 gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag
    15661 cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc
    15721 ttgggggttc cagtgccatt gcagggccgg caggcaaccc agccgcttac gcctggccaa
    15781 ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt
    15841 tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta
    15901 ctctggtagc tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac
    15961 cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat
    16021 ggctggcgtg tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg
    16081 gccggcactt agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg
    16141 agttttgatt taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt
    16201 tctgattcaa gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg
    16261 atacgggact caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg
    16321 atgcgcgtgc ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg
    16381 acctcaatgc gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg
    16441 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
    16501 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
    16561 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
    16621 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
    16681 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
    16741 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
    16801 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta
    16861 gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat
    16921 cagacaaacc ggccaggatt tcatgcagcc gcacggttga gacgtgcgcg ggcggctcga
    16981 acacgtaccc ggccgcgatc atctccgcct cgatctcttc ggtaatgaaa aacggttcgt
    17041 cctggccgtc ctggtgcggt ttcatgcttg ttcctcttgg cgttcattct cggcggccgc
    17101 cagggcgtcg gcctcggtca atgcgtcctc acggaaggca ccgcgccgcc tggcctcggt
    17161 gggcgtcact tcctcgctgc gctcaagtgc gcggtacagg gtcgagcgat gcacgccaag
    17221 cagtgcagcc gcctctttca cggtgcggcc ttcctggtcg atcagctcgc gggcgtgcgc
    17281 gatctgtgcc ggggtgaggg tagggcgggg gccaaacttc acgcctcggg ccttggcggc
    17341 ctcgcgcccg ctccgggtgc ggtcgatgat tagggaacgc tcgaactcgg caatgccggc
    17401 gaacacggtc aacaccatgc ggccggccgg cgtggtggtg tcggcccacg gctctgccag
    17461 gctacgcagg cccgcgccgg cctcctggat gcgctcggca atgtccagta ggtcgcgggt
    17521 gctgcgggcc aggcggtcta gcctggtcac tgtcacaacg tcgccagggc gtaggtggtc
    17581 aagcatcctg gccagctccg ggcggtcgcg cctggtgccg gtgatcttct cggaaaacag
    17641 cttggtgcag ccggccgcgt gcagttcggc ccgttggttg gtcaagtcct ggtcgtcggt
    17701 gctgacgcgg gcatagccca gcaggccagc ggcggcgctc ttgttcatgg cgtaatgtct
    17761 ccggttctag tcgcaagtat tctactttat gcgactaaaa cacgcgacaa gaaaacgcca
    17821 ggaaaagggc agggcggcag cctgtcgcgt aacttaggac ttgtgcgaca tgtcgttttc
    17881 agaagacggc tgcactgaac gtcagaagcc gactgcacta tagcagcgga ggggttggat
    17941 caaagtactt tgatcccgag gggaaccctg tggttggcat gcacatacaa atggacgaac
    18001 ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct
    18061 tag
//
