LOCUS       sequence-487277-       18804 bp    DNA     circular SYN 26-SEP-2025
DEFINITION  Gateway-based vector for Marchantia polymorpha to express a floxed 
            gene, with an expression cassette for Cre-glucocorticoid receptor 
            fusion protein under the control of heat-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18804)
  AUTHORS   Nishihama R, Ishida S, Urawa H, Kamei Y, Kohchi T
  TITLE     Conditional Gene Expression/Deletion Systems for Marchantia 
            polymorpha Using its Own Heat-shock Promoter and the 
            Cre/loxP-mediated Site-specific Recombination.
  JOURNAL   Plant Cell Physiol. 2015 Jul 6. pii: pcv102.
  PUBMED    26148498
REFERENCE   2  (bases 1 to 18804)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell 
            Physiol. 2015 Jul 6. pii: pcv102."
COMMENT     Sequence Label: pMpGWB337
FEATURES             Location/Qualifiers
     source          1..18804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    54..78
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     266..288
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     280..297
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2042..2075)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2089..2213
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2250..2255,2256..2273,2274..2280)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2334..2993
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2400..2419)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3335..3640
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3519..3538
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3681..3805)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3823..4075
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4108..4124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4132..4148
                     /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(4156..4162,4163..4180,4181..4186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4212..4245)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(4251..4253,4254..4256,4257..4967)
                     /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(4557..4576)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4904..4925
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4974..4994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5019..5271
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5352..5368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5376..5392
                     /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(5400..5406,5407..5424,5425..5430))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5445..5466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    6698..6722
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6743..7771
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     protein_bind    complement(7788..7812)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      8714..8966
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8999..9015)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9023..9039
                     /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(9047..9053,9054..9071,9072..9077))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9092..9113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    9198..9372
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(11455..12131)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(11498..11518)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    12326..12350
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             12871..13662
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      13908..14496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14397..14416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14650..14667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    14682..14822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(14748..14767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14908..14930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      15166..15360
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(15426..16499)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(16928..17557)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
       61 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
      121 tccaagctca agctgctcta gcattcgcca ttcaggctgc gcaactgttg ggaagggcga
      181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      301 aagcttgggt ctagaatgag tcacacacat tgttgagaga catatcaaaa taactcaatt
      361 gtaaggtaag ttgatagatt tcccagttaa atgcattatt ttaacaatat ttgtatctca
      421 aattttcaaa atcatatcaa atttgcagat cttgatgatg attcacaact ctgtcttcga
      481 tttttgattg ttaacgggta tcgatatttg ggtaacttaa aaagaatttg ctaaagatct
      541 taaaaaacag attaaaaaaa cctttcaaaa tagctaatcc tttaaatccg tttcacaata
      601 taatgctcat tgatgataat cgaaaagatt ccattttaaa gtgcatataa attgcttaat
      661 tatctctttc tgtctcgtaa tattttgttc tctcctattg gatcggaccc gcatcttctc
      721 gatcagttca ctatactaat atttggtcaa cttcaagaga gattagaact tcgttctcca
      781 aagtgtaaag gccgacaagc agaaaaggaa acttcaatca gaagccttct aggttttccc
      841 gcgtcccaaa aagcgaaggg aggacgggga ggacggtgag gccagaagaa gaaagcaaca
      901 gtaaaatctc atgtgattgc gtgcatgtca tggatctatg ggatgctagt ccaccgtttg
      961 gaagtatgtg cagaaggaga aaggcgcggg aggccaaaag cgacttcggc cacctctcta
     1021 ctttctagaa agtttgctcg cagcttgtaa taagcgcgga aaaggtactc gaaaaaccgg
     1081 agcaatcggt gtggtatcag cagtcgggtt cgctaaacct acgtcttctt attttggaat
     1141 gttaaattga aagggcgtgc cggccctaag cacggacgat gtgacagacg gaggagtgtg
     1201 agcaattcgg atgacgatgc tggccggcag tcacgacaaa gagtccgaag gggcatggac
     1261 agggcaatat ataaatggaa aagaggcttt gaatattcct aacccttcgt cgtcggcctc
     1321 ccaacttgcc ctcgcatgct gaattggccg cctatctgcg aatggccggc cgccgaccct
     1381 aagtcgcaat aacttgaacg gagccgatgc aaagtcctta tcgaacgctc gttattataa
     1441 acggggcaga cctaggtatt taattatatc aaaaccctcc gcaatttcgc taatcattcc
     1501 ctaatttctt ctactttggc gtctagtgct gtggtcgcgc ttgctctcaa agctcgctgg
     1561 gaggcagtgg agcgtctggc ttaaaatcga tcgccgcaac cctagagttc ctttcacttc
     1621 tttctctctt gcgtggagag catcgacatc gaagagctgc atcttgtctt ctggtatgcg
     1681 ccttgtgccc tcgtctctgt acgtcgttac gattgctcga tattgtgatt gtctagacct
     1741 gaaacccgac tggagctttt acgtggtgtc gattggggcc tcgagcgagt gattttttag
     1801 gttcgtttta gccaagtaca tcccactttg gtcagtcctg ttatctcggt aaggtgttca
     1861 gttcggtctc tggtgcggag gtgcctcgta agtttcctgc atgatagatt tgtctatcct
     1921 tgacttgaag aggagtcgaa ggtcagcgag gttggctgtg tgctatggac gtcggtgctc
     1981 cttggttccg tgcttctatg ggtgttgcct caacagtatt gatgtgcctg cagaaaggtt
     2041 gataacttcg tatagcatac attatacgaa gttattctag agttatcaac aagtttgtac
     2101 aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     2161 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggcggccc
     2221 ctcgaggggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
     2281 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
     2341 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     2401 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     2461 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     2521 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     2581 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     2641 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     2701 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2761 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2821 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     2881 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2941 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
     3001 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     3061 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     3121 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     3181 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     3241 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     3301 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     3361 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     3421 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     3481 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     3541 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     3601 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     3661 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     3721 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     3781 gttcagcttt cttgtacaaa gtggttgata acagcgctta gcgatcgttc aaacatttgg
     3841 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt
     3901 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     3961 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     4021 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa
     4081 ttggttccgg aaccaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     4141 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4201 atgagtgagc tataacttcg tatagcatac attatacgaa gttatccacc atggtgagca
     4261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4321 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4381 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4441 ccttcggcta cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact
     4501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4681 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     4741 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     4801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     4861 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagttg cagcctaaga
     4981 agaagagaaa ggttggagga tagagctcga atttccccga tcgttcaaac atttggcaat
     5041 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     5101 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     5161 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     5221 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattgg
     5281 ttccggaacc aattggttcc ggaaccaatt ggttccggaa ccaattggtt ccggaaccaa
     5341 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5401 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5461 acattaggct gaattaggcg cgcagccatt atagcaacta tgttaaaagt gacagttttc
     5521 tctaatcggg aatcttgtcc gtattgaaag ttccctgttc agtgactgag ccgcttaata
     5581 atattttcag tttcgagtac gagggagaga ttgaacaaat tttggggttt cagggctaaa
     5641 ttctgcaggg gctttagttt tgagtcgggt gaaactttcc taagtcttgg caatgaactc
     5701 acattcgcca ttgtacctat taaatgaccc aaacatcctt gacaacaaat tgacaagtac
     5761 gggaacatga tcttcaatat gaaatttaat atgtccttta tgttggcttt cctcatcaat
     5821 gattggaaac tcacatacaa agaaaagcgt gatataataa atcactagaa tcttctccgc
     5881 tttctaagac agtaatgatg gatccacgtg agtgtgaaac atcaatatca acatcggtag
     5941 gagtagagaa gtttctaaat cagaaaaagt gaatagacaa gacaacttac tccttgaacg
     6001 agaatattcc tcgagaaaga gggaaaagag taaaaagagt aaaaagccat ggtacagaat
     6061 agcccattcc atggttattg cagtcaaacc gctacgccag caaaaacgga acgaaactaa
     6121 acccgttcca caaattgcga aagatggaat gtattcgcca tgtatatttc gagattatta
     6181 tgagacttcc ggtcccggga agatggcctg ggttcaccgg gatcagccga attccgacgt
     6241 gatggagcac ggccatgcat acttctagag aagtcggcca ccgtgaaatt cgacgacgag
     6301 atggtactag aatcaccggg acagtacaca cctcgttcat ggcctcgagc attcgatgtc
     6361 gctctcacct gcacagcctc gaggtatcga gagggttcta ggatggacca gcgatgtctc
     6421 gaggaattta gaaagatttc atgcgctcgt tgtgtgagag ccaaggaagc acattatggg
     6481 gcctataagt accacgcaag ggcctggggt tcccatcaat caaatcagtg ttcagacgat
     6541 ttcctcaagc gatcattgca ccgtgatcag tgaagcacag acttcaagct tttggcttgc
     6601 ctacttccac gttcccatcc gagatcgaca ccgaattgtt gagttctgtc tgatcttcct
     6661 gcagtcgaga gcgaagttcc accatctaga gttatcaaca agtttgtaca aaaaagcagg
     6721 ctccgcggcc gcccccttca ccatgtccaa tttactgacc gtacaccaaa atttgcctgc
     6781 attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga
     6841 tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc
     6901 ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga
     6961 ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc aacatttggg
     7021 ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt
     7081 ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca
     7141 ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga
     7201 tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg
     7261 tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat
     7321 gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact
     7381 tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga
     7441 tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac
     7501 cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta
     7561 cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt
     7621 cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg
     7681 tggctggacc aatgtaaata ttgtcatgaa ctatatccgt aacctggata gtgaaacagg
     7741 ggcaatggtg cgcctgctgg aagatggcga taagggtggg cgcgccgacc cagctttctt
     7801 gtacaaagtg gttgataaca gcatggctag tgaagctcga aaaacaaaga aaaaaatcaa
     7861 agggattcag caagccactg caggagtctc acaagacact tcggaaaatc ctaacaaaac
     7921 aatagttcct gcagcattac cacagctcac ccctaccttg gtgtcactgc tggaggtgat
     7981 tgaacccgag gtgttgtatg caggatatga tagctctgtt ccagattcag catggagaat
     8041 tatgaccaca ctcaacatgt taggtgggcg tcaagtgatt gcagcagtga aatgggcaaa
     8101 ggcgatacca ggcttcagaa acttacacct ggatgaccaa atgaccctgc tacagtactc
     8161 atggatgttt ctcatggcat ttgccctggg ttggagatca tacagacaat caagtggaaa
     8221 cctgctctgc tttgctcctg atctgattat taatgagcag agaatgtctc taccctgcat
     8281 gtatgaccaa tgtaaacaca tgctgtttgt ctcctctgaa ttacaaagat tgcaggtatc
     8341 ctatgaagag tatctctgta tgaaaacctt actgcttctc tcctcagttc ctaaggaagg
     8401 tctgaagagc caagagttat ttgatgagat tcgaatgact tatatcaaag agctaggaaa
     8461 agccatcgtc aaaagggaag ggaactccag tcagaactgg caacggtttt accaactgac
     8521 aaaacttctg gactccatgc atgaggtggt tgagaatctc cttacctact gcttccagac
     8581 atttttggat aagaccatga gtattgaatt cccagagatg ttagctgaaa tcatcactaa
     8641 tcagatacca aaatattcaa atggaaatat caagaagctg ctgtttcatc aaaaatagag
     8701 ctcgaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt
     8761 tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat
     8821 taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt
     8881 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     8941 cgcggtgtca tctatgttac tagatcggga attggttccg gaaccaattc gtaatcatgt
     9001 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     9061 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaggctgaa
     9121 ttaggcgcgc ctatttctga attcaacaca ttgcggacgt ttttaatgta ctgaattaac
     9181 gccgaattaa ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa
     9241 ttttattgat agaagtattt tacaaataca aatacatact aagggtttct tatatgctca
     9301 acacatgagc gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt
     9361 attatggaga aactcgagtt agtatatagt ccttccatca ccctctgtga taatctcctt
     9421 aaaacttgct cccccaggaa tcataggcag aacgtgctcc tggtggggaa ctatcacatc
     9481 cagtaaataa ggaccgggag aatctagcat agtttgaatt gctgctctta gttcatcctt
     9541 cttcaccaca cgcgcatttg gcaccttgaa accgtctgca actgtacaga agtcggggta
     9601 gatctgctcc tcgtgatctg gatcacctaa gtaagtatgt gctctgttgg ccttgtagaa
     9661 tctatcctcc aattgtacga ccattcctag atgttggttg tttaagatca tgatcttcac
     9721 gggtaacttc tccacaaaaa ttgtagccaa ctcctgtacg ttcatcacaa agctaccatc
     9781 gccatcgatg tcgatcacag gaatatctgg ttgagttgcg gcagccccca aagcagaagg
     9841 aagtccgaat cccatggctc caagacctcc agatgtcaac caacgccgcg gttcgttgta
     9901 cttgaaccac tgggctgccc acatttgatg ttggccaaca ccagtagata tgatggcatt
     9961 tccaccagtg agttcgtaca gcgtctgtat ggcagattga ggagcaatgg catcctcgaa
    10021 tacgggaaac ttcattggcc atttctgctt ttgttcttct agttctgcca cccagttgga
    10081 gaagtcgaaa gtcctctttt gactcttcaa gagggcattg atttcactta gcgcaggttt
    10141 cacatcagca cacatggaca catggggctg tttgttcttt ccaatttcag cagggtcaat
    10201 atcgatatga acgatgctgg acctgctagc aaaagtctcc agtttaccag tcactcgatc
    10261 gtcgaaacgc actccaaatg ccaacaacag atctgcttgg tccacggcgt agttggcgta
    10321 aacagttcca tgcattccaa gcattttcaa acttaatgga tcagactcgg gatacgctcc
    10381 caaacccatt aaagtcgatg tgataggaat tccggtaagc tcgacaaatt ccctcaactc
    10441 ttcgctggcg tgattgcaac ctcctccaga atacactaca ggcttcttag aactagaaat
    10501 gagtcttata atctgctcga ttaacgttgg ttccggtggc cgaggcagtc gttgtaagta
    10561 gcccttcagt tgcattggct gagaccaatc tggaatagcc atttgctgtt gcacgtcctt
    10621 gggaatatct accaagactg ggcccggcct acctgaagct gccaaaaaga aagcctccct
    10681 gattacgcga gggatgtcat caatgttcat aacaaggtaa ttgtgcttgg tgatcgatct
    10741 cgtaacctcc acaattggag tttcttgaaa agcatcggta ccaatcatcc tactggatac
    10801 ttgacctgta atggccacta gtggcacact gtccaagagg gcatcagcga gcccggtaac
    10861 aagattggtc gctccaggac cggatgtggc tatacaaacc cctactcgtc cagacgattt
    10921 agcataacct tcagcagcga aaatctcccc ctgctcgtgg cgacagagaa tgttgcggat
    10981 gctgctggaa cgtgtaagag cctggtgaat ctccatagac gctccgccgg gataagcgaa
    11041 caccgttcta atcccttctc gctcaagagc ctccaccaaa atgtcggctc ctttgcgggg
    11101 ctcgtcatcc ttgtagcgag actgaaagtc ggacggggtt ggtgcaacga gtgacttccc
    11161 gctgctaagc cgtggtccac tcacatcgac tgcagactgg atcttctcgc taacagcttg
    11221 ggctttcact tccagtgaac gctcagtctt ccgagaggtg ttgtatgcat gtcccctcag
    11281 agcggctccc gagaagaaac cactcctcac ctttagctct ttagctccct tgcaacaaga
    11341 ccttgccgat ggctgcacct tctccaggct cgctatgctc aaaggcacag ctgcgcatcc
    11401 actaattgac gccatgctcg agagagatag atttgtagag agagactggt gatttcagcg
    11461 tgtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg
    11521 ggattgtgcg tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga
    11581 cgtggttgga acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg
    11641 accactgtcg gcagaggcat cttgaacgat agcctttcct ttatcgcaat tatggcattt
    11701 gtaggtgcca ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg
    11761 gaatccgagg aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc
    11821 ttctgagact gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt
    11881 cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc
    11941 tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct
    12001 ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga
    12061 tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt
    12121 gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga
    12181 cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgaattc
    12241 agaaataaat tcagcctaat tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt
    12301 tattaagttg tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc
    12361 aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag
    12421 tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga
    12481 tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag
    12541 ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcaat tcgggcacga
    12601 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca
    12661 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
    12721 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
    12781 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
    12841 ggcagtcgcc ctaaaacaaa gttaaacatc atgggggaag cggtgatcgc cgaagtatcg
    12901 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
    12961 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
    13021 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
    13081 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
    13141 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
    13201 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
    13261 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
    13321 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
    13381 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
    13441 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
    13501 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
    13561 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
    13621 cactacgtga aaggcgagat caccaaggta gtcggcaaat aatgtctagc tagaaattcg
    13681 ttcaagccga cgccgcttcg cggcgcggct taactcaagc gttagatgca ctaagcacat
    13741 aattgctcac agccaaacta tcaggtcaag tctgctttta ttatttttaa gcgtgcataa
    13801 taagccctac acaaattggg agatatatca tgcatgacca aaatccctta acgtgagttt
    13861 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
    13921 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
    13981 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
    14041 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
    14101 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
    14161 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
    14221 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
    14281 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
    14341 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
    14401 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
    14461 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
    14521 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
    14581 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
    14641 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
    14701 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
    14761 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
    14821 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
    14881 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
    14941 catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac
    15001 ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg
    15061 ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg
    15121 gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg
    15181 cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc
    15241 tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca
    15301 atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca
    15361 ggaaagagac cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc
    15421 gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg
    15481 atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg
    15541 atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg
    15601 tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg
    15661 tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg
    15721 tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg
    15781 tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc
    15841 tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc
    15901 gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct
    15961 cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct
    16021 cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc
    16081 aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg
    16141 tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa
    16201 acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg
    16261 aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc
    16321 ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg
    16381 cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc
    16441 aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgctcacg
    16501 ccgattcctc gggcttgggg gttccagtgc cattgcaggg ccggcagaca acccagccgc
    16561 ttacgcctgg ccaaccgccc gttcctccac acatggggca ttccacggcg tcggtgcctg
    16621 gttgttcttg attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt
    16681 catttgctca tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc
    16741 ttgccttggc gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag
    16801 ttgacccgct tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg
    16861 cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct
    16921 cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag
    16981 cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc
    17041 tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg
    17101 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta
    17161 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc
    17221 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg
    17281 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga
    17341 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat
    17401 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa
    17461 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg
    17521 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt
    17581 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac
    17641 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc
    17701 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ttgagacgtg
    17761 cgcgggcggc tcgaacacgt acccggccgc gatcatctcc gcctcgatct cttcggtaat
    17821 gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg cttgttcctc ttggcgttca
    17881 ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt cctcacggaa ggcaccgcgc
    17941 cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa gtgcgcggta cagggtcgag
    18001 cgatgcacgc caagcagtgc agccgcctct ttcacggtgc ggccttcctg gtcgatcagc
    18061 tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc gggggccaaa cttcacgcct
    18121 cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga tgattaggga acgctcgaac
    18181 tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg ccggcgtggt ggtgtcggcc
    18241 cacggctctg ccaggctacg caggcccgcg ccggcctcct ggatgcgctc ggcaatgtcc
    18301 agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg tcactgtcac aacgtcgcca
    18361 gggcgtaggt ggtcaagcat cctggccagc tccgggcggt cgcgcctggt gccggtgatc
    18421 ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt cggcccgttg gttggtcaag
    18481 tcctggtcgt cggtgctgac gcgggcatag cccagcaggc cagcggcggc gctcttgttc
    18541 atggcgtaat gtctccggtt ctagtcgcaa gtattctact ttatgcgact aaaacacgcg
    18601 acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc gcgtaactta ggacttgtgc
    18661 gacatgtcgt tttcagaaga cggctgcact gaacgtcaga agccgactgc actatagcag
    18721 cggaggggtt ggatcaaagt actttgatcc cgaggggaac cctgtggttg gcatgcacat
    18781 acaaatggac gaacggataa acct
//
